阅读说明：本技术 负载维生素d的柠檬酸钙/硫酸钙复合骨水泥及制备方法 (Vitamin D-loaded calcium citrate/calcium sulfate composite bone cement and preparation method thereof ) 是由 严永刚 邓光进 陈红 戢觅之 于 2019-11-25 设计创作，主要内容包括：本发明属于骨修复材料领域,具体涉及一种载维生素D的柠檬酸钙/硫酸钙复合骨水泥及制备方法与应用。本发明提供一种负载维生素D的柠檬酸钙/硫酸钙复合骨水泥,所述骨水泥由固相部分和液相部分组成,液固比为0.2～0.6：1；固相部分包括柠檬酸钙、硫酸钙盐和维生素D,柠檬酸钙和硫酸钙盐的质量比为1：4～4：1；维生素D与骨水泥的质量比为6×10<Sup>-3</Sup>～20×10<Sup>-3</Sup>：1。本发明所得复合骨水泥具有良好的可注射性、合适的凝固时间、缓慢的药物释放和优异的生物活性,可以用于骨缺损的填充使用。(The invention belongs to the field of bone repair materials, and particularly relates to vitamin D-loaded calcium citrate/calcium sulfate composite bone cement, and a preparation method and application thereof. The invention provides vitamin D-loaded calcium citrate/calcium sulfate composite bone cement, which consists of a solid-phase part and a liquid-phase part, wherein the liquid-solid ratio is 0.2-0.6: 1; the solid phase part comprises calcium citrate, calcium sulfate salt and vitamin D, wherein the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 4-4: 1; the mass ratio of the vitamin D to the bone cement is 6 multiplied by 10 ‑3 ～20×10 ‑3 : 1. the composite bone cement prepared by the invention has good injectability, proper setting time, slow drug release and excellent bioactivity, and can be used for filling bone defects.)

1. The vitamin D-loaded calcium citrate/calcium sulfate composite bone cement is characterized by comprising a solid-phase part and a liquid-phase part, wherein the liquid-solid ratio is 0.2-0.6: 1; the solid phase part comprises calcium citrate, calcium sulfate salt and vitamin D, wherein the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 4-4: 1; the mass ratio of the vitamin D to the bone cement is 6 multiplied by 10^-3～20×10^-3：1。

2. The vitamin D-loaded calcium citrate/calcium sulfate composite bone cement as claimed in claim 1, wherein the calcium sulfate salt is at least one of α -calcium sulfate hemihydrate, β -calcium sulfate hemihydrate or calcium sulfate dihydrate.

3. The vitamin D-loaded calcium citrate/calcium sulfate composite bone cement according to claim 1 or 2, wherein the calcium citrate is at least one of calcium citrate tetrahydrate, calcium citrate dihydrate or anhydrous calcium citrate.

4. The vitamin D-loaded calcium citrate/calcium sulfate composite bone cement as claimed in any one of claims 1 to 3, wherein the solid phase part further comprises at least one of sodium alginate, hyaluronic acid or gelatin.

5. The calcium citrate/calcium sulfate composite bone cement loaded with vitamin D as claimed in any one of claims 1 to 4, wherein the vitamin D is at least one of vitamin D2, vitamin D3, 1, 25-dihydroxy vitamin D2 or 1, 25-dihydroxy vitamin D3.

6. The preparation method of the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement as claimed in any one of claims 1 to 5, wherein the preparation method comprises the following steps:

1) uniformly mixing calcium citrate and calcium sulfate salt to form a solid-phase compound, wherein the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 4-4: 1;

2) uniformly stirring the solid phase compound and a vitamin D-ethanol solution, and then drying and crushing to obtain solid phase blend powder; wherein the concentration of the vitamin D-ethanol solution is 0.1-12.5 mg/mL, and the liquid-solid ratio of the vitamin D-ethanol solution to the solid phase compound is 1.5-3 mL/g;

3) mixing and stirring the solid-phase blend powder and the curing liquid to form bone cement paste, and then forming to obtain the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement; wherein the liquid-solid ratio of the curing liquid to the solid-phase blend is 0.2-0.3 mL/g.

7. The method for preparing the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement according to claim 6, wherein in the step 1), the mass ratio of the calcium citrate to the calcium sulfate is 2: 3-3: 2.

8. the preparation method of the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement according to claim 6 or 7, characterized in that in the step 1), calcium citrate and calcium sulfate are uniformly mixed in a ball milling manner, and the ball milling time is 2-12 hours; the ball milling speed is 130-170 r/min.

9. The method for preparing the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement according to any one of claims 6 to 8, wherein in the step 2), the particle size of the solid-phase blend is up to 150 meshes, and the solid-phase blend is sieved.

10. The method for preparing the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement according to any one of claims 6 to 9, wherein in the step 2), the concentration of the vitamin D-ethanol solution is 0.1 to 2.5 mg/mL.

Technical Field

The invention belongs to the field of bone repair materials, and particularly relates to vitamin D-loaded calcium citrate/calcium sulfate composite bone cement, and a preparation method and application thereof.

Background

In recent years, with the aging of the population, the incidence of various wounds and chronic diseases such as osteoporosis has been remarkably increased, and how to effectively treat osteoporosis has become one of the hot problems of research. Calcium is the main mineral element that constructs human bones. Calcium not only contributes to bone formation, but also activates various enzymes. 99% of calcium in human body is stored in hard tissues such as bones and teeth, and only 1% of calcium exists in soft tissues such as blood, viscera, muscles and nerves in an ionic state. Calcium deficiency can cause rickets, osteomalacia and osteoporosis, and can also cause hypertension, arteriosclerosis, soft tissue calcium, senile dementia and other diseases. In addition, research shows that vitamin D has a certain prevention effect on osteoporosis. Therefore, for osteoporosis patients, it is necessary to fill bone graft materials during early healing of bone fracture and release sufficient calcium ions and vitamin D to bone tissues to help bone tissue healing.

Currently, clinically used bone filling materials include hydroxyapatite, calcium phosphate, calcium silicate, calcium sulfate, etc., but each of them has some problems, such as low strength, too fast or too slow degradation, poor injectability, etc., which limits its clinical application. Therefore, there is a need to develop a new bone repair material to better meet clinical needs.

Disclosure of Invention

The invention aims to provide the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement which has good injectability, proper setting time, slow drug release and excellent bioactivity and can be used for filling bone defects.

The technical scheme of the invention is as follows:

the invention aims to solve the first technical problem of providing calcium citrate/calcium sulfate composite bone cement loaded with vitamin D, wherein the bone cement consists of a solid phase part and a liquid phase part, and the liquid-solid ratio is 0.2-0.6: 1; the solid phase part comprises calcium citrate, calcium sulfate salt and vitamin D, wherein the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 4 to 4: 1; the mass ratio of the vitamin D to the bone cement is 6 multiplied by 10^-3～20×10^-3：1。

Further, the calcium sulfate salt is at least one of α -calcium sulfate hemihydrate, β -calcium sulfate hemihydrate or calcium sulfate dihydrate.

Further, the calcium citrate is calcium citrate tetrahydrate Ca₃(C₆H₅O₇)₂·4H₂O, calcium citrate dihydrate Ca₃(C₆H₅O₇)₂·2H₂O or anhydrous calcium citrate Ca₃(C₆H₅O₇)₂At least one of (1).

Further, the solid phase part also comprises at least one of sodium alginate, hyaluronic acid or gelatin. The compressive strength and the formability of the bone cement can be improved by adding a proper amount of sodium alginate, hyaluronic acid or gelatin.

Further, the vitamin D is at least one of vitamin D2 (calciferol), vitamin D3 (cholecalciferol ), 1, 25-dihydroxyvitamin D2, or 1, 25-dihydroxyvitamin D3.

The second technical problem to be solved by the invention is to provide a preparation method of the calcium citrate/calcium sulfate composite bone cement loaded with vitamin D, which comprises the following steps:

1) uniformly mixing calcium citrate and calcium sulfate salt to form a solid-phase compound, wherein the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 4-4: 1;

2) uniformly stirring the solid phase compound and a vitamin D-ethanol solution, and then drying and crushing to obtain solid phase blend powder; wherein the concentration of the vitamin D-ethanol solution is 0.1-12.5 mg/mL, and the liquid-solid ratio of the vitamin D-ethanol solution to the solid phase compound is 1.5-3 mL/g;

3) mixing and stirring the solid-phase blend powder and the curing liquid to form bone cement paste, and then forming to obtain the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement; wherein the liquid-solid ratio of the curing liquid to the solid-phase blend is 0.2-0.3 mL/g.

Further, in the step 1), the mass ratio of calcium citrate to calcium sulfate is 2: 3-3: 2.

further, in the step 1), calcium citrate and calcium sulfate are uniformly mixed in a ball milling mode, wherein the ball milling time is 2-12 hours; preferably 4 to 8 hours, and the ball milling speed is 130 to 170 revolutions per minute.

Further, in the step 2), the particle size of the solid-phase blend reaches 150 meshes and is sieved.

Preferably, in the step 2), the concentration of the vitamin D-ethanol solution is 0.1-2.5 mg/mL.

The invention has the beneficial effects that:

the invention firstly compounds calcium citrate and calcium sulfate salt into the form of bone cement, and quantitatively and stably loads a proper amount of vitamin D, so that the prepared bone cement has slow drug release and good bone formation capability, and the calcium sulfate salt is added to obviously improve the collapsibility resistance of the calcium citrate and ensure good plasticity. The vitamin D-ethanol solution with different concentrations is added into the composite bone cement, the physical and chemical properties of the composite bone cement are not changed, but the cell activity is obviously improved. The calcium citrate/calcium sulfate composite bone cement loaded with vitamin D can be widely applied to a bone defect method due to proper setting time, good injectability, higher mechanical strength and slow drug release.

Description of the drawings:

FIG. 1 is a graph showing the setting time of bone cements obtained in examples 1 to 6.

FIG. 2 is a graph showing injectability of bone cements obtained in examples 1-6.

FIG. 3 is a graph showing the compressive strengths of bone cements obtained in examples 1 to 6.

FIG. 4 is an XRD diffractogram of bone cements obtained in examples 1-6.

FIG. 5 is a graph showing the setting time of vitamin D-loaded bone cements obtained in examples 7 to 11.

Fig. 6 is a graph of injectability of vitamin D-loaded bone cements of examples 7-11.

FIG. 7 is a graph of the compressive strength of vitamin D-loaded bone cements of examples 7-11.

FIG. 8 is the degradation curves of vitamin D-loaded bone cements of examples 7-11 in PBS solution for 42 days.

FIG. 9 is the drug release profile of vitamin D loaded bone cements of examples 7-11 in PBS solution for 42 days.

Detailed Description

The invention aims to solve the first technical problem of providing calcium citrate/calcium sulfate composite bone cement loaded with vitamin D, wherein the bone cement consists of a solid phase part and a liquid phase part, and the liquid-solid ratio is 0.2-0.6: 1; wherein the solid phase part is formed by compounding calcium citrate and calcium sulfate salt, and the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 9-9: 1; the mass ratio of the vitamin D to the bone cement is 0-20 x 10^-3：1。

In the invention, calcium citrate can release citrate ions in the hydration process, and COO in the calcium citrate^-The functional group can be chelated with calcium ions in the calcium sulfate to form a coordination bond, so that water is prevented from entering the calcium sulfate, the curing process of the calcium sulfate is slowed down, and the setting time of the composite bone cement is obviously longer than that of the calcium sulfate.

The second technical problem to be solved by the invention is to provide a preparation method of the calcium citrate/calcium sulfate composite bone cement loaded with vitamin D, which comprises the following steps:

1) uniformly mixing calcium citrate and calcium sulfate salt to form a solid-phase compound, wherein the mass ratio of the calcium citrate to the calcium sulfate salt is 1: 9-9: 1;

2) uniformly stirring the solid phase compound and a vitamin D-ethanol solution, and then drying and crushing to obtain solid phase blend powder; wherein the concentration of the vitamin D-ethanol solution is 0.1-12.5 mg/mL, and the liquid-solid ratio of the vitamin D-ethanol solution to the solid phase compound is 1.5-3 mL/g;

3) mixing and stirring the solid-phase blend powder and the curing liquid to form bone cement paste, and then forming to obtain the vitamin D-loaded calcium citrate/calcium sulfate composite bone cement; wherein the liquid-solid ratio of the curing liquid to the solid-phase blend is 0.2-0.3 mL/g; 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.

In the invention, a vitamin D-ethanol solution is required, otherwise, the vitamin D cannot be uniformly dispersed into the bone cement, and the stable calcium citrate/calcium sulfate composite bone cement carrying the vitamin D cannot be formed; in addition, the vitamin D content in the bone cement is less than the minimum value of the invention, and the cell proliferation promoting function is not performed, and if the vitamin D content is more than the maximum value of the invention, the vitamin poisoning phenomenon may occur.

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.