CN111803708A - 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating and preparation method and application thereof - Google Patents
3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of biomedical materials, and discloses a 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating, and a preparation method and application thereof. The preparation method comprises the following steps: 1) preparing a gelatin/sodium alginate/hydroxyapatite composite scaffold by a 3D printing technology, sequentially crosslinking with calcium chloride and genipin, soaking with L-sodium glutamate, washing with deionized water in the middle, and soaking for multiple times; (2) dissolving dopamine hydrochloride, curcumin macromolecular drug and gelatin in Tris-HCl with the pH value of 8.5 by stirring; (3) immersing the bracket into the solution obtained immediately, vibrating at constant temperature, taking out and quickly freezing for crosslinking; (4) and (3) soaking the scaffold in L-sodium glutamate after the genipin is crosslinked again, washing the scaffold with deionized water for many times, and obtaining the curcumin macromolecule/polydopamine synergistic anticancer coating on the surface of the hydroxyapatite scaffold.
Description
Technical Field
The invention belongs to the technical field of biomedical materials, and particularly relates to a 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating, and a preparation method and application thereof.
Background
Osteosarcoma is the most common primary malignant bone tumor in orthopedics department, the onset age mainly has 2 peaks, the first peak is <24 years old, and the incidence rate is 4.4 per 100 ten thousand years; the second peak is >59 years, with a 4.2/100 million years of morbidity, with the prominent sites of good hair being the femur, tibia and humerus. The clinical treatment means for osteosarcoma mainly comprises surgical treatment, radiotherapy/chemotherapy and interventional treatment. The traditional solution for surgical treatment is amputation, but this solution usually causes serious limb dysfunction to the patient, and the long-term survival rate (>5 years) of the patient can only reach 20%. Therefore, in recent years, the main choice of clinical surgery is limb protection surgery, which can preserve the limb function of the patient to the maximum extent and improve the life quality of the patient, but the clinical surgery may increase the recurrence rate of osteosarcoma and is extremely dependent on experienced operators. Traditional radiotherapy/chemotherapy treatment can help patients to protect limbs to the maximum extent and improve the survival quality, but the traditional radiotherapy/chemotherapy has great side effects, and can only improve the long-term survival rate to 70-75% under the best condition, and the survival rate cannot be achieved generally. Interventional therapy comprises microwave ablation, radio frequency ablation, argon-helium knife ablation and the like, has the advantages of small damage, less pain and high safety factor due to treatment under image guidance, but has the same risk of possibly increasing the recurrence rate as limb protection surgery, the long-term survival rate of recurrent osteosarcoma can only reach about 20%, and complications can be induced.
At present, the clinical osteosarcoma treatment mainly adopts a scheme of limb protection surgery and radiotherapy/chemotherapy assistance, but the surgery can cause bone defects which cannot be cured by self, and simultaneously, the complete excision of all tumor cells is difficult to ensure, and the traditional radiotherapy/chemotherapy can bring serious side effects. Therefore, it is of great significance to design a biomaterial which can kill possible residual tumor cells while having little influence on normal cells and is beneficial to bone tissue repair after killing the residual tumor cells.
Curcumin (Curcumin, Cur) is a polyphenol substance extracted from rhizome of plant belonging to genus Curcuma of family Zingiberaceae. Researches show that the curcumin has wide pharmacological activity and can efficiently generate ROS under certain illumination conditions; toxicological studies have demonstrated that it remains safe and reliable at high doses, and has been classified as a third-generation cancer chemopreventive by the FDA. But the curcumin has extremely poor solubility in water, is unstable under physiological conditions and is metabolized quickly in vivo, so that the bioavailability is low, and the clinical application is greatly limited.
Photothermal therapy has received much attention because of its non-invasive, effective and non-toxic side effects. Polydopamine (PDA) as a melanin analogue has strong near-infrared absorption capacity and high photothermal conversion efficiency, and unlike other widely used photothermal materials, PDA has excellent biocompatibility and biodegradability. Photothermal therapy kills cancer cells at high temperature and also has a great effect on surrounding normal tissues, while below 43 ℃ cancer cells are only inhibited in activity. In addition, studies have shown that dopamine may be addictive and not be used in excess.
The dual-mode tumor synergistic treatment has high efficiency, drug resistance and lower side effect, so that the dual-mode tumor synergistic treatment becomes a tumor treatment mode which is concerned in recent years. Compared with the traditional tumor treatment mode, the photodynamic therapy and the photothermal therapy are more accurate and efficient, and have no wound and low toxic and side effects. Under the condition of reasonable design, the photodynamic therapy and the photothermal therapy can be simultaneously started under one illumination condition, so that the photodynamic therapy and the photothermal therapy are synergistically acted on tumor tissues, the treatment means is greatly simplified, and the tumor treatment efficiency is improved. Meanwhile, the invention adopts a macromolecular curcumin drug obtained by modifying polyethylene glycol, compared with a single curcumin drug, the water solubility and the stability of the drug are greatly improved, the metabolic rate is improved, and the anticancer activity is kept.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention mainly aims to provide a preparation method of a 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating.
The invention also aims to provide the 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating prepared by the preparation method.
The invention further aims to provide application of the 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating.
The purpose of the invention is realized by the following technical scheme:
a preparation method of a 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating comprises the following steps:
(1) preparing a gelatin/sodium alginate/hydroxyapatite composite scaffold by a 3D printing technology, quickly crosslinking for 15min by using 5 wt% of calcium chloride, washing by using deionized water, crosslinking for 72h at room temperature in a dark place by using 0.5 wt% of genipin, taking out, washing again by using the deionized water, soaking for 48h by using 0.5 wt% of L-sodium glutamate, and then soaking for 24h by using the deionized water;
(2) dissolving dopamine hydrochloride in a Tris-hydrochloric acid solution (Tris-HCl) with the pH value of 8.5, then sequentially adding a curcumin macromolecular drug (pCur) and gelatin, and fully stirring and dissolving in a water bath kettle at the temperature of 40 ℃ to obtain a mixed solution, wherein the curcumin macromolecular drug has the following structural formula:
n1is a natural number of 42 to 50, n2Is a natural number more than or equal to 5;
(3) immersing the hydroxyapatite scaffold treated in the step (1) into the mixed solution obtained in the step (2) for constant-temperature oscillation, taking out, and immediately placing into a refrigerator at minus 80 ℃ for quick freezing and crosslinking;
(4) and (3) taking out the quickly-frozen and crosslinked stent in the step (3), crosslinking the quickly-frozen and crosslinked stent for 12h at 4 ℃ in a dark place by 0.5 wt% of genipin, washing the stent by deionized water, soaking the stent by 0.5 wt% of L-sodium glutamate for 2h, and washing the stent by the deionized water to obtain the curcumin macromolecule/polydopamine synergistic anticancer coating on the surface of the hydroxyapatite stent, so as to obtain the 3D-printed hydroxyapatite stent composite curcumin macromolecule/polydopamine synergistic anticancer coating.
The preparation of the gelatin/sodium alginate/hydroxyapatite composite scaffold by the 3D printing technology in the step (1) specifically comprises the following steps: dissolving diammonium phosphate in deionized water, and adjusting the pH value of the solution to 6.0 by using nitric acid to obtain a diammonium phosphate solution; dissolving calcium nitrate hexahydrate in deionized water to obtain a calcium nitrate solution; mixing a diammonium hydrogen phosphate solution and a calcium nitrate solution, adding sodium citrate, stirring for 15min, transferring to a high-pressure reaction kettle, reacting for 3h at 180 ℃ to obtain hydroxyapatite microsphere powder, washing with deionized water, aging, and freeze-drying for later use; sequentially adding sodium alginate, gelatin and hydroxyapatite microsphere powder into ultrapure water, heating and stirring in a water bath to obtain uniform slurry, transferring into a charging barrel, ultrasonically removing all bubbles, and preparing the gelatin/sodium alginate/hydroxyapatite composite scaffold by using a 3D printing technology.
The curcumin macromolecular drug in the step (2) is prepared according to the following steps: reacting polyethylene glycol, succinic anhydride and p-toluenesulfonic acid together at 80 ℃ in a nitrogen environment; dissolving the product obtained by the reaction in a solvent, filtering, precipitating with a precipitator, centrifuging, dissolving again with the solvent, repeatedly purifying for 3 times to obtain a product, freeze-drying, adding the product, curcumin and dicyclohexylcarbodiimide into the solvent, fully dissolving, charging nitrogen, adding 4-dimethylaminopyridine and triethylamine dissolved with the solvent, stirring for reaction for 24 hours, precipitating with the precipitator after filtering, centrifuging, precipitating again with the precipitator after dissolving with the solvent, repeatedly purifying for 3 times, and freeze-drying to obtain a curcumin macromolecule; the solvent is dichloromethane, and the precipitant is anhydrous diethyl ether.
The preparation method of the tris (hydroxymethyl) aminomethane-hydrochloric acid solution with the pH of 8.5 in the step (2) comprises the following steps: 0.61g of tris (hydroxymethyl) aminomethane-hydrochloric acid is weighed and dissolved in 500mL of water, after complete dissolution, the pH value is measured by a pH meter, and 0.5M hydrochloric acid solution is dropwise added until the pH value is adjusted to 8.5.
The final concentration of the dopamine hydrochloride in the mixed solution in the step (2) is 0.2-0.4mg/mL, the final concentration of the curcumin macromolecular drug in the mixed solution is 1-2mg/mL, and the final content of the gelatin in the mixed solution is 5 wt%;
the constant-temperature shaking temperature in the step (3) is 40 ℃, and the time is 30 min; the quick-freezing crosslinking time is 30 min;
and (5) pre-freezing the 3D printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating overnight, and then freeze-drying to obtain a freeze-dried sample.
The step of lyophilizing is followed by a step of aseptic processing.
In each step, water used for preparing all the solutions is laboratory self-made ultrapure water.
The 3D printing hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating prepared by the preparation method.
The 3D printed hydroxyapatite scaffold is applied to a repair material for a bone defect part after osteosarcoma operation by compounding the curcumin macromolecule/polydopamine synergistic anti-cancer coating.
Compared with the prior art, the invention has the following advantages and effects:
the invention designs a hydroxyapatite 3D printing support with a pCur/PDA composite coating, wherein the synthesized curcumin macromolecules have good water solubility and physiological stability, the composite coating locally and relatively low promotes the temperature of a tumor microenvironment by utilizing a small amount of PDA under a certain illumination condition, and the composite coating is cooperated with ROS (reactive oxygen species) generated by the curcumin macromolecules, so that the influence on surrounding normal tissues is reduced to the maximum degree, and meanwhile, possibly residual osteosarcoma cells are killed; the coating part can start photodynamic therapy and photothermal therapy while being rapidly degraded, so that the coating part can act on tumor tissues in a synergistic manner, the treatment means is greatly simplified, the tumor treatment efficiency is improved, and the hydroxyapatite scaffold part exposed by the degradation of the coating is favorable for repairing the bone defect part after osteosarcoma operation.
Drawings
FIG. 1 is the NMR spectrum of curcumin macromolecular drug pCur.
Fig. 2 is a physical diagram of a 3D-printed hydroxyapatite scaffold compounded with curcumin macromolecules/polydopamine synergistic anticancer coating, and it can be seen that an obvious coating is formed on the surface of the scaffold.
Fig. 3 is a surface Scanning Electron Microscope (SEM) and energy spectrum element analysis diagram of the 3D printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating; the SEM figure illustrates that the coating is coated on the 3D scaffold and on the surface with holes that form the asperities; meanwhile, the C, O, N element distribution in the energy spectrum is matched with the SEM image, which shows that pCur and PDA can exist in the coating uniformly.
Fig. 4 is a longitudinal section Scanning Electron Microscope (SEM) image of the 3D printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating, with magnifications of 1000 times, 500 times, 100 times and 50 times, respectively; in the longitudinal section SEM image, a loose porous coating part and a compact stent part are obviously arranged, the coating part uniformly wraps the stent part, and meanwhile, the two parts can be tightly combined as seen in a 1000-time enlarged image.
Fig. 5 is a cross-sectional Scanning Electron Microscope (SEM) and energy spectrum elemental analysis (spectroscopy) image of a 3D printed hydroxyapatite scaffold compounded curcumin macromolecule/polydopamine synergistic anticancer coating; SEM images also show that the loosely porous coating structure can be loaded evenly into the internal structure of the dense 3D scaffold section. Meanwhile, P, Ca elements in the energy spectrum only exist in the structural formula of the scaffold containing hydroxyapatite, and C, O, N elements are uniformly distributed, which indicates that the coating loaded into the direct interior contains pCur and PDA.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1:
the hydroxyapatite powder is self-made in a laboratory, is a microsphere with a micro-nano structure, has an average particle size of 10 mu m, and the preparation method comprises the following steps:
dissolving diammonium phosphate in deionized water to obtain a solution with the final concentration of 24mmol/L, and adjusting the pH of the solution to 6 by using nitric acid; dissolving calcium nitrate hexahydrate in deionized water to obtain a solution with the final concentration of 40 mmol/L; mixing the two solutions, adding 3.6g of sodium citrate, stirring vigorously for 15min, transferring into a high-pressure reaction kettle with a polytetrafluoroethylene lining, reacting at 180 ℃ for 3h, washing with ethanol and deionized water, aging, and freeze-drying to obtain hydroxyapatite powder for later use.
Example 2:
the preparation method of the gelatin/sodium alginate/hydroxyapatite composite scaffold comprises the following steps:
(1) preparing printing slurry: sequentially adding 4 wt% of sodium alginate, 10 wt% of gelatin and 10 wt% of the hydroxyapatite powder prepared in the embodiment 1 into ultrapure water, stirring uniformly in a 50 ℃ water bath, quickly transferring to a 3D printing material cylinder, performing ultrasonic treatment at 40 ℃ until no bubbles exist, and quickly cooling in a refrigerator at-20 ℃ for 10min for later use;
(2) 3D printing of the composite support: modeling by utilizing software design, obtaining a cylindrical model file with the overall size of phi 10 multiplied by 2mm, the layer height of 0.32mm and 90-degree rotation between layers, guiding the cylindrical model file into a 3D printer, setting the temperature of a charging barrel to be 30-32 ℃, the temperature of a receiving table to be 0 ℃, the extrusion pressure parameter to be 2.5-3 bar and the printing speed to be 8-10 mm/s, and printing by a 0.4mm needle head at the ambient temperature of 24 ℃ to obtain an uncrosslinked gelatin/sodium alginate/hydroxyapatite composite bracket with the internal fiber diameter of 0.4-0.6 mm and the same-layer fiber spacing of 0.4-0.6 mm;
(3) crosslinking of the composite scaffold: and (3) rapidly crosslinking the printed gelatin/sodium alginate/hydroxyapatite composite scaffold for 15min by using 5 wt% of calcium chloride, washing by using deionized water, crosslinking for 72h at room temperature in a dark place by using 0.5 wt% of genipin, taking out, washing again by using deionized water, soaking for 48h by using 0.5 wt% of L-sodium glutamate, soaking for 24h by using deionized water to obtain the crosslinked gelatin/sodium alginate/hydroxyapatite composite scaffold, and freeze-drying and storing for later use.
Example 3:
curcumin macromolecules (pCur) are self-made in laboratories, the molecular weight is about 20000, and the curcumin drug loading rate is about 14.6%. Polyethylene glycol in the synthesis raw materials was purchased from Sigma and had a number average molecular weight of 2050. The preparation method comprises the following steps:
(1) adding 8g of polyethylene glycol with the number average molecular weight of 2050 into a flask, sequentially adding 1.17g of succinic anhydride and 0.332g of p-toluenesulfonic acid, and reacting for 3 hours at 80 ℃ in a nitrogen environment;
(2) dissolving the product obtained after the reaction in 20ml of dichloromethane, filtering, precipitating the product by 160ml of anhydrous ether, centrifuging at room temperature, dissolving again by 20ml of dichloromethane, repeatedly purifying for 3 times, pre-freezing overnight and freeze-drying the obtained product;
(3) dissolving 750mg of curcumin, 4.58g of the product obtained in the step (2) and 1.26g of DCC into 150ml of ultra-dry dichloromethane, fully dissolving, charging nitrogen, adding 0.1g of DMAP and 0.1ml of TEA dissolved by the ultra-dry dichloromethane, and reacting at room temperature for 24h under vigorous stirring;
(4) filtering the reacted solution, vacuumizing and concentrating the solution to half of the original volume, precipitating with anhydrous ether with the volume ratio of 8 times, dissolving with dichloromethane at room temperature, precipitating with anhydrous ether again, and repeating the operation for 3 times to obtain the curcumin macromolecular drug pCur;
(5) the resulting pCur drug was prefrozen overnight and lyophilized for use. The nuclear magnetic resonance hydrogen spectrum is shown in figure 1, and the appearance of characteristic peaks indicates the success of the synthesis.
The obtained curcumin macromolecular drug has the following structural formula:
n1is a natural number of 42 to 50, n2Is a natural number more than or equal to 5.
Example 4:
a preparation method of a 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating comprises the following steps:
(1) weighing 0.61g of Tris, dissolving in 500mL of water, measuring the pH value of the solution by using a pH meter after the solution is completely dissolved, and dropwise adding 0.5M hydrochloric acid solution until the pH value is adjusted to 8.5;
(2) weighing dopamine hydrochloride, dissolving the dopamine hydrochloride in Tris-HCl (pH 8.5) prepared in the step (1), sequentially adding curcumin macromolecular drug pCur and gelatin obtained in the example 3, and fully stirring and dissolving the curcumin macromolecular drug pCur and the gelatin in a water bath kettle at 40 ℃ to obtain a mixed solution, wherein the final concentration of the dopamine hydrochloride solution is 0.2mg/mL, the final concentration of the pCur solution is 1mg/mL, and the final content of the gelatin is 5 wt%; (ii) a
(3) Putting the cross-linked gelatin/sodium alginate/hydroxyapatite composite scaffold obtained in the example 2 into a 24-pore plate, adding 2ml of the mixed solution obtained in the step (2) into each pore, shaking at the constant temperature of 40 ℃ for 30min, taking out, and immediately putting into a refrigerator at the temperature of-80 ℃ for quick freezing and cross-linking for 30 min;
(4) taking out the quick-frozen and crosslinked stent, crosslinking the quick-frozen and crosslinked stent for 12 hours at 4 ℃ in a dark place by 0.5 wt% of genipin, soaking the quick-frozen and crosslinked stent for 2 hours by 0.5 wt% of L-sodium glutamate at normal temperature after washing by deionized water, and then washing by the deionized water; thus obtaining the pCur/PDA synergistic anticancer coating on the surface of the hydroxyapatite scaffold;
(5) the hydroxyapatite scaffold compounded with the pCur/PDA synergistic anticancer coating is subjected to pre-freezing overnight, freeze-drying and gamma ray irradiation sterilization treatment to obtain the 3D printed hydroxyapatite scaffold compounded with the curcumin macromolecules/polydopamine synergistic anticancer coating.
Example 5:
in the steps (1), (3), (4) and (5) of the preparation method of the synergistic anticancer coating, like in example 4, in the step (2), dopamine hydrochloride is weighed and dissolved in freshly prepared Tris-HCl (pH 8.5), then pCur and gelatin are added in sequence, and the mixture is fully stirred and dissolved in a water bath kettle at 40 ℃, wherein the final concentration of the dopamine hydrochloride solution is 0.4mg/mL, the final concentration of the pCur solution is 1mg/mL, and the final content of the gelatin is 5 wt%.
Example 6:
in the steps (1), (3), (4) and (5) of the preparation method of the synergistic anticancer coating, like in example 4, in the step (2), dopamine hydrochloride is weighed and dissolved in freshly prepared Tris-HCl (pH 8.5), then pCur and gelatin are added in sequence, and the mixture is fully stirred and dissolved in a water bath kettle at 40 ℃, wherein the final concentration of the dopamine hydrochloride solution is 0.2mg/mL, the final concentration of the pCur solution is 2mg/mL, and the final content of the gelatin is 5 wt%.
Example 7:
in the steps (1), (3), (4) and (5) of the preparation method of the synergistic anticancer coating, like in example 4, in the step (2), dopamine hydrochloride is weighed and dissolved in freshly prepared Tris-HCl (pH 8.5), then pCur and gelatin are added in sequence, and the mixture is fully stirred and dissolved in a water bath kettle at 40 ℃, wherein the final concentration of the dopamine hydrochloride solution is 0.4mg/mL, the final concentration of the pCur solution is 2mg/mL, and the final content of the gelatin is 5 wt%.
Example 8:
the 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating obtained in example 4 was photographed by a camera in a wet state, and the results are shown in fig. 2. After freeze drying, the scaffold is cut from the middle by a blade to obtain samples of the surface, the cross section and the longitudinal section, the samples are fixed in an aluminum sample table by conductive adhesive, gold spraying treatment is carried out for 90 seconds, observation is carried out by a scanning electron microscope, and energy spectrum element analysis is carried out, and the result is shown in fig. 3-5.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of a 3D-printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating is characterized by comprising the following steps:
(1) preparing a gelatin/sodium alginate/hydroxyapatite composite scaffold by a 3D printing technology, quickly crosslinking for 15min by using 5 wt% of calcium chloride, washing by using deionized water, crosslinking for 72h at room temperature in a dark place by using 0.5 wt% of genipin, taking out, washing again by using the deionized water, soaking for 48h by using 0.5 wt% of L-sodium glutamate, and then soaking for 24h by using the deionized water;
(2) dissolving dopamine hydrochloride in a trihydroxymethyl aminomethane-hydrochloric acid solution with the pH value of 8.5, sequentially adding curcumin macromolecular drugs and gelatin, and fully stirring and dissolving in a water bath kettle at the temperature of 40 ℃ to obtain a mixed solution; the curcumin macromolecular drug has the following structural formula:
n1is a natural number of 42 to 50, n2Is a natural number more than or equal to 5;
(3) immersing the hydroxyapatite scaffold treated in the step (1) into the mixed solution obtained in the step (2) for constant-temperature oscillation, taking out, and immediately placing into a refrigerator at minus 80 ℃ for quick freezing and crosslinking;
(4) and (3) taking out the quickly-frozen and crosslinked stent in the step (3), crosslinking the quickly-frozen and crosslinked stent for 12h at 4 ℃ in a dark place by 0.5 wt% of genipin, washing the stent by deionized water, soaking the stent by 0.5 wt% of L-sodium glutamate for 2h, and washing the stent by the deionized water to obtain the curcumin macromolecule/polydopamine synergistic anticancer coating on the surface of the hydroxyapatite stent, so as to obtain the 3D-printed hydroxyapatite stent composite curcumin macromolecule/polydopamine synergistic anticancer coating.
2. The method of claim 1, wherein: the preparation of the gelatin/sodium alginate/hydroxyapatite composite scaffold by the 3D printing technology in the step (1) specifically comprises the following steps: dissolving diammonium phosphate in deionized water, and adjusting the pH value of the solution to 6.0 by using nitric acid to obtain a diammonium phosphate solution; dissolving calcium nitrate hexahydrate in deionized water to obtain a calcium nitrate solution; mixing a diammonium hydrogen phosphate solution and a calcium nitrate solution, adding sodium citrate, stirring for 15min, transferring to a high-pressure reaction kettle, reacting for 3h at 180 ℃ to obtain hydroxyapatite microsphere powder, washing with deionized water, aging, and freeze-drying for later use; sequentially adding sodium alginate, gelatin and hydroxyapatite microsphere powder into ultrapure water, heating and stirring in a water bath to obtain uniform slurry, transferring into a charging barrel, ultrasonically removing all bubbles, and preparing the gelatin/sodium alginate/hydroxyapatite composite scaffold by using a 3D printing technology.
3. The method of claim 1, wherein: the curcumin macromolecular drug in the step (2) is prepared according to the following steps: reacting polyethylene glycol, succinic anhydride and p-toluenesulfonic acid together at 80 ℃ in a nitrogen environment; dissolving the product obtained by the reaction in a solvent, filtering, precipitating with a precipitator, centrifuging, dissolving again with the solvent, repeatedly purifying for 3 times to obtain a product, freeze-drying, adding the product, curcumin and dicyclohexylcarbodiimide into the solvent, fully dissolving, charging nitrogen, adding 4-dimethylaminopyridine and triethylamine dissolved with the solvent, stirring for reaction for 24 hours, precipitating with the precipitator after filtering, centrifuging, precipitating again with the precipitator after dissolving with the solvent, repeatedly purifying for 3 times, and freeze-drying to obtain a curcumin macromolecule; the solvent is dichloromethane, and the precipitant is anhydrous diethyl ether.
4. The method of claim 1, wherein: the preparation method of the tris (hydroxymethyl) aminomethane-hydrochloric acid solution with the pH of 8.5 in the step (2) comprises the following steps: 0.61g of tris (hydroxymethyl) aminomethane-hydrochloric acid is weighed and dissolved in 500mL of water, after complete dissolution, the pH value is measured by a pH meter, and 0.5M hydrochloric acid solution is dropwise added until the pH value is adjusted to 8.5.
5. The method of claim 1, wherein: the final concentration of the dopamine hydrochloride in the mixed solution in the step (2) is 0.2-0.4mg/mL, the final concentration of the curcumin macromolecular drug in the mixed solution is 1-2mg/mL, and the final content of the gelatin in the mixed solution is 5 wt%.
6. The method of claim 1, wherein: the constant-temperature shaking temperature in the step (3) is 40 ℃, and the time is 30 min; the time for quick-freezing and crosslinking is 30 min.
7. The method of claim 1, wherein: and (5) pre-freezing the 3D printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating overnight, and then freeze-drying to obtain a freeze-dried sample.
8. The method of claim 7, wherein: the step of lyophilizing is followed by a step of aseptic processing.
9. A3D printed hydroxyapatite scaffold composite curcumin macromolecule/polydopamine synergistic anticancer coating prepared by the preparation method of any one of claims 1 to 8.
10. The 3D-printed hydroxyapatite scaffold compounded curcumin macromolecule/polydopamine synergistic anti-cancer coating according to claim 9 is applied to a repair material for a bone defect part after osteosarcoma operation.
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