CN109529109B

CN109529109B - Preparation method of overlong-structure metal ion-doped hydroxyapatite fiber material

Info

Publication number: CN109529109B
Application number: CN201811247733.XA
Authority: CN
Inventors: 杨辉; 叶射稳; 伍阳君; 蒋赞丽; 杨佳琦
Original assignee: Jiangxi University of Science and Technology
Current assignee: Jiangxi University of Science and Technology
Priority date: 2018-10-25
Filing date: 2018-10-25
Publication date: 2021-01-05
Anticipated expiration: 2038-10-25
Also published as: CN109529109A

Abstract

Discloses a preparation method of a metal ion doped hydroxyapatite fiber material with an ultra-long structure, which comprises the following steps: (1) mixing the ion-doped salt, the soluble calcium salt, guar gum and disodium hydrogen phosphate with deionized water to obtain a first mixed solution; (2) injecting the first mixed solution into a strong base solution to form fiber gel, continuously soaking the obtained fiber gel in the strong base solution for a certain time, and then drying to obtain a fiber precursor; (3) and calcining the fiber precursor to obtain a product, washing the product with deionized water, and drying to obtain the ultra-long structure metal ion doped hydroxyapatite fiber material. The method has the advantages of simple preparation process, easily controlled conditions and no need of any template agent.

Description

Preparation method of overlong-structure metal ion-doped hydroxyapatite fiber material

Technical Field

The invention belongs to the field of preparation of biomedical materials, and particularly relates to a preparation method of a metal ion doped hydroxyapatite fiber material with an ultra-long structure.

Background

Hydroxyapatite (HAp) has the characteristics of degradability, good biocompatibility, osteoconductivity, certain osteoinduction and the like, so that the hydroxyapatite has a very wide application prospect in the field of bone repair. The hydroxyapatite fiber material has the advantages of easy molding, large specific surface area and the like compared with other materials, but the prior hydroxyapatite fiber material has relatively complex synthetic process and very low yield; in addition, the construction method of the hydroxyapatite porous scaffold is generally divided into three steps: the process of synthesizing, forming and calcining the material is relatively long. In conclusion, the development of a method for preparing hydroxyapatite fiber materials in large batch is a difficult problem in the related field at present when the construction time of a porous scaffold is shortened.

During the repair of bone defects, post-operative infection occurs around the implant due to the possible adhesion of bacteria to the surface of the material. Researches show that the antibacterial performance of the hydroxyapatite ceramic material can be improved by a copper doping mode; in addition, the metal ion doping can also obviously enhance the bone repair process of the hydroxyapatite ceramic material and promote the generation of new bone tissues; meanwhile, researches show that the manganese-doped hydroxyapatite can promote the repair of hard tissues and has very important influence on the repair process of cartilage tissues.

Disclosure of Invention

The invention provides a preparation method of a super-long structure metal ion doped hydroxyapatite fiber material, which comprises the following steps:

(1) mixing the ion-doped salt, the soluble calcium salt, guar gum and disodium hydrogen phosphate with deionized water to obtain a first mixed solution;

(2) injecting the first mixed solution obtained in the step (1) into a strong alkali solution to form fiber gel, continuously soaking the obtained fiber gel in the strong alkali solution for a certain time, and then drying to obtain a fiber precursor;

(3) and (3) calcining the fiber precursor obtained in the step (2) to obtain a product, washing the product with deionized water, and drying to obtain the ultra-long structure metal ion doped hydroxyapatite fiber material.

In the present invention, "salt of a doping ion" means a corresponding salt of a metal ion desired to be doped in a material. For example, if a manganese ion doped hydroxyapatite fibrous material is desired, the salt is a manganese salt (e.g., in example 2, the salt of the doping ion is manganese nitrate, ultimately obtaining an ultralong structure Mn ion doped hydroxyapatite fibrous material). Further, in the step (1), the salt of the dopant ion may be a salt selected from magnesium, strontium, barium, manganese, cobalt, iron, nickel, copper, and zinc; soluble salts are preferred, and may be chlorides, nitrates and acetates.

In the present invention, the "soluble calcium salt" may be calcium nitrate, calcium chloride or other calcium salt capable of forming an aqueous solution.

Further, in the step (1), the amount of each substance used may be such that a solution is prepared according to the following criteria: in the obtained first mixed solution, the concentration of soluble calcium salt is 0.005-1 mol/L; the concentration of the disodium hydrogen phosphate is 0.005-1 mol/L; the concentration of the guar gum is 0.1-10 g/L. The concentration of the salt of the dopant ion is determined in accordance with practical use, and experiments prove that when the concentration is more than 0 and less than or equal to 0.1mol/L, the addition of the salt of the dopant ion has no substantial influence on the molding of the material.

Further, in the step (1), the specific preparation process of the first mixed solution may be: (a) dissolving soluble calcium salt, ion-doped salt and guar gum in deionized water, stirring until the soluble calcium salt, the ion-doped salt and the guar gum are completely dissolved, then adding disodium hydrogen phosphate, and stirring until the soluble calcium salt, the ion-doped salt and the guar gum are completely dissolved. It can also be: (b) soluble calcium salt, ion-doped salt, and disodium hydrogen phosphate are dissolved in deionized water, stirred to completely dissolve, followed by addition of guar gum, and stirring to completely dissolve. It will be understood that the speed and duration of agitation is not limited, subject to agitation to complete homogeneity. In some embodiments, the two steps can be stirred at 10-2000 rpm for 2-24 hours.

Further, the soluble calcium salt and the salt doped with ions used in the present invention may be in the form of a salt with or without crystal water (for example, in some embodiments, the soluble calcium salt used is calcium nitrate tetrahydrate, Ca (NO)₃)₂·4H₂O, the salt used as a doping ion is anhydrous copper chloride CuCl₂). Guar gum for use in the present invention may be in a form including, but not limited to, powder; adding guar gumIn the preparation method, the guar gum powder can be uniformly added by using a screen while stirring.

Further, in the present invention, the first mixed solution is injected into the strong alkali solution, and a method that may be adopted includes, but is not limited to, injecting the first mixed solution into the strong alkali solution using a syringe. It is understood that the injection speed and the injection hole diameter are not limited to enable the first mixed solution to be stably injected into the strong alkali solution and to obtain a fiber gel of a proper size; in some embodiments, the injection rate may be 1-10 mL/s; experiments prove that the method is suitable for conventional injection pore sizes.

Further, in the step (2), the strong alkali solution can be a sodium hydroxide solution or a potassium hydroxide solution, and the concentration can be 0.1-10 mol/L; the soaking time is 1-240 hours.

Further, in the step (2), the volume ratio of the first mixed solution to the strong alkali solution may be 1:10 to 10: 1.

Further, in the step (2), the drying may be freeze drying or oven drying. In some embodiments, the drying process may be performed at a drying temperature of 40 to 120 ℃ for 2 to 72 hours. In other embodiments, the drying process may be carried out at-80 to-20 ℃ (further, preferably at-60 to-20 ℃), and the vacuum drying is carried out for 1 to 2 days (as will be understood by those skilled in the art, pre-cooling is further required before the vacuum drying is carried out).

In step (3), the fiber precursor is calcined to obtain a product, and as will be understood by those skilled in the art, the calcination method includes, but is not limited to: the calcination is carried out in a high temperature furnace. Further, the calcination may be performed in an air atmosphere, and the process conditions may be: the heating rate is 1-20 ℃/min, the temperature is increased from room temperature to 800-1250 ℃, the temperature is preserved for 1-6 hours, and then the product is naturally cooled to room temperature.

In step (3), the product is washed with deionized water as a conventional procedure, and the effect thereof includes removing soluble impurities in the product, washing the product to neutrality, and the like, as will be understood by those skilled in the art. One specific operation is: soaking and cleaning for 1-48 hours each time, and repeating for a plurality of times. Another specific operation is: directly washed with a large amount of deionized water.

In the step (3), the product is washed with water and then dried, which is a conventional operation. In certain embodiments, the process conditions for drying in step (3) may be: the drying temperature is room temperature to 300 ℃, and the drying time is 1 to 72 hours.

The invention also aims to provide a super-long structure metal ion doped hydroxyapatite fiber material prepared by the preparation method. The fiber material is a porous structure, and the diameter of the material can be 100nm to 500 mu m according to the pore diameter of an injector. In some embodiments, the fiber material prepared by the above method may have a length of tens of centimeters or even tens of centimeters.

It is another object of the present invention to provide the use of the above materials in bone tissue repair.

In general, the obtained fibrous material is in a loose form. However, if the fibrous material is transferred to a container (e.g., a test tube) having a small space (relative to the length of the fiber) before drying in step (2), then the resulting fibrous material is not unraveled but entangled to form a porous scaffold material (i.e., to form a shaped scaffold material) after drying and calcination due to the limited stretching of the gel fibers. Therefore, the invention also provides a hydroxyapatite bracket which is composed of the fiber material. The fiber material in the form of a scaffold is prepared by the method, and the step (2) can be freeze drying or oven drying. The scaffold formed by freeze-drying will have a greater porosity than the two drying methods. Namely, the hydroxyapatite porous scaffold prepared by the method has simple process and easily controlled conditions.

The preparation method of the ultra-long structure metal ion doped hydroxyapatite fiber material has the following advantages:

(1) the preparation process of the system is simple, the conditions are easy to control, and no template agent is needed. The size, the composition and the gel behavior of the ultra-long structure metal ion doped hydroxyapatite fiber material can be regulated by adjusting various parameters (including but not limited to pinhole size, metal ion concentration and Ca/P ratio).

(2) The ultra-long structure metal ion doped hydroxyapatite fiber material prepared by the system has good biocompatibility and capability of promoting bone cell proliferation.

(3) Only water is required as a solvent in the preparation process.

According to the invention, the hydroxyapatite composite gel fiber is obtained by one-step method by utilizing the strong physical crosslinking action between guar gum and alkaline earth or transition metal ions in an alkaline solution system, then the gel is dried to obtain a metal ion doped hydroxyapatite fiber precursor material, and the metal ion doped hydroxyapatite fiber can be directly obtained after the precursor material is calcined, so that the construction process of the hydroxyapatite composite scaffold system is greatly shortened, and a new thought is provided for the construction of a novel hydroxyapatite scaffold.

In the prior art, hydroxyapatite fibers can be generally prepared by a method such as electrospinning, however, the existing method for preparing hydroxyapatite fibers with a super-long structure (tens of centimeters to tens of centimeters) has several defects. At present, the electrostatic spinning method is the most common method for preparing fiber materials, but the method has the following defects: high pressure environment, high requirement on equipment, long preparation period, low efficiency and difficulty in forming a three-dimensional structure. On the contrary, the preparation method is simple, the first mixed solution can be injected into the strong alkali solution under the conditions of normal temperature and normal pressure to form the fiber gel under the condition of not using a template agent, and the three-dimensional porous scaffold structure can be obtained after the fiber gel is dried.

Drawings

Fig. 1 and fig. 2 are optical images of a precursor of an ultra-long structure metal ion-doped hydroxyapatite fiber material according to embodiment 1 of the present invention;

fig. 3 is an XRD of the ultra-long structure Cu-doped hydroxyapatite fiber material of example 1 of the present invention;

fig. 4 is an XRD of the Mn-doped hydroxyapatite fiber material with an ultra-long structure of example 2 of the present invention.

Fig. 5 is a porous scaffold prepared from the Mn-doped hydroxyapatite fiber material with an ultra-long structure according to embodiment 9 of the present invention.

Fig. 6 shows the influence of the overlong Mn-doped hydroxyapatite fiber material according to embodiment 9 on the proliferation behavior of the mesenchymal stem cells.

Detailed Description

The invention is further illustrated by the following examples.

Example 1

(1) Firstly, taking CuCl₂、Ca(NO₃)₂·4H₂O、Na₂HPO₄The solid was added to deionized water and stirred at 200rpm for 2 hours to give a homogeneous solution. Then weighing guar gum powder, adding guar gum powder into the solution with a screen while stirring, stirring at 200rpm for 3 hr to obtain 200mL first mixed solution containing CuCl₂Is 0.0001 mol/L, Ca (NO)₃)₂·4H₂The concentration of O is 0.05mol/L, the concentration of disodium hydrogen phosphate is 0.03mol/L, and the concentration of guar gum is 5 g/L.

(2) The first mixed solution was injected into 100mL of a 1mol/L potassium hydroxide solution at a rate of 5mL/s using a syringe (inner diameter of a needle: 0.2mm, the same applies hereinafter) to form gel fibers, and the solution was maintained for further 24 hours. Then dried at 50 ℃ for 6 hours to obtain a fiber precursor.

(3) And putting the fiber precursor into a crucible, transferring the crucible into a high-temperature furnace, heating to 800 ℃ at a heating rate of 5 ℃/min in the air atmosphere, preserving the heat for 2 hours, and naturally cooling to room temperature to obtain the product. And soaking the calcined product in deionized water for 24 hours (repeating for 3 times), then placing the calcined product in an oven for 6 hours at 50 ℃, and drying to obtain the overlong structure Cu-doped hydroxyapatite fiber material with the length of about 20-40 cm and the diameter of about 0.2 mm.

Example 2

(1) Firstly, Mn (NO) is taken₃)₂、Ca(NO₃)₂·4H₂O、Na₂HPO₄The solid was added to deionized water and stirred at 200rpm for 2 hoursA homogeneous solution is obtained. Then weighing guar gum powder, adding guar gum powder into the solution with a screen while stirring, stirring at 200rpm for 3 hr to obtain 200mL of first mixed solution containing Mn (NO)₃)₂Has a concentration of 0.0007mol/L, Ca (NO)₃)₂·4H₂The concentration of O is 0.5mol/L, the concentration of disodium hydrogen phosphate is 0.3mol/L, and the concentration of guar gum is 5 g/L.

(2) The first mixed solution was injected into 100mL of 1mol/L NaOH solution at a rate of 5mL/s by a syringe to form gel fibers, and the solution was maintained for further 72 hours. Then dried at 100 ℃ for 2 hours to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 1000 ℃ at the heating rate of 20 ℃/min under the air atmosphere, preserving the heat for 1 hour, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying for 48 hours at 50 ℃, and obtaining the overlong structure Mn ion doped hydroxyapatite fiber material after drying, wherein the length is about 30-50 cm, and the diameter is about 0.2 mm.

Example 3

(1) Taking Zn (NO) first₃)₂、CaCl₂、Na₂HPO₄The solid was added to deionized water and stirred at 100rpm for 2 hours to give a homogeneous solution. Then weighing guar gum powder, adding guar gum powder into the solution with a screen while stirring, stirring at 1000rpm for 2 hr to obtain 200mL first mixed solution, wherein Zn (NO) is added₃)₂The concentration of (1) is 0.004mol/L, CaCl₂The concentration of the compound is 0.05mol/L, the concentration of disodium hydrogen phosphate is 0.03mol/L, and the concentration of guar gum is 1 g/L.

(2) The first mixed solution was injected into 100mL of 10mol/L NaOH solution at a rate of 10mL/s by a syringe to form gel fibers, and the solution was maintained for further 2 hours. Then dried at 120 ℃ for 2 hours to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 800 ℃ at a heating rate of 1 ℃/min under the air atmosphere, preserving the heat for 6 hours, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying for 3 hours at 100 ℃, and obtaining the Zn ion doped hydroxyapatite fiber material with the ultra-long structure after drying, wherein the length is about 20-30 cm, and the diameter is about 0.2 mm.

Example 4

(1) Firstly taking Ni (CH)₃COO)₂、Ca(NO₃)₂·4H₂O、Na₂HPO₄The solid was added to deionized water and stirred at 10rpm for 24 hours to give a homogeneous solution. Then weighing guar gum powder, adding guar gum powder into the solution with a screen while stirring, stirring at 300rpm for 4 hr to obtain 200mL of first mixed solution containing Ni (CH)₃COO)₂Has a concentration of 0.0097mol/L, Ca (NO)₃)₂·4H₂The concentration of O is 0.05mol/L, the concentration of disodium hydrogen phosphate is 0.03mol/L, and the concentration of guar gum is 3 g/L.

(2) The first mixed solution was injected into 400mL of 7mol/L NaOH solution at a rate of 8mL/s by a syringe to form gel fibers, and the solution was maintained for another 10 hours. Then freezing for 5 hours at the temperature of 60 ℃ below zero, and then drying for 2 days in vacuum to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 1250 ℃ at the heating rate of 10 ℃/min in the air atmosphere, preserving the heat for 1 hour, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying at 300 ℃ for 1 hour to obtain the Ni ion doped hydroxyapatite fiber material with the ultra-long structure, wherein the length is about 50 cm, and the diameter is about 0.2 mm.

Example 5

(1) Taking Ni (NO) first₃)₂、Ca(NO₃)₂·4H₂O、Na₂HPO₄Adding the solid into deionized water, and stirring at 10rpm for 24 hours to obtain a uniform solutionAnd (4) homogenizing the solution. Then weighing guar gum powder, adding guar gum powder into the solution with a screen mesh while stirring, stirring at 300rpm for 2 hr to obtain 1000mL of first mixed solution, wherein Ni (NO) is added₃)₂Has a concentration of 0.0000065mol/L, Ca (NO)₃)₂·4H₂The concentration of O is 0.005mol/L, the concentration of disodium hydrogen phosphate is 0.005mol/L, and the concentration of guar gum is 0.1 g/L.

(2) The first mixed solution was injected into 100mL of 0.1mol/L NaOH solution at a rate of 1mL/s by a syringe to form gel fibers, and the solution was maintained for a further 240 hours. Then dried at 120 ℃ for 2 hours to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 1100 ℃ at a heating rate of 15 ℃/min under the air atmosphere, preserving the heat for 1.5 hours, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying for 72 hours at 50 ℃, and obtaining the Ni ion doped hydroxyapatite fiber material with an ultra-long structure after drying, wherein the length is about 40-50 cm, and the diameter is about 0.2 mm.

Example 6

(1) Taking Co (CH) first₃COO)₂、Ca(NO₃)₂·4H₂O、Na₂HPO₄The solid was added to deionized water and stirred at 50rpm for 12 hours to give a homogeneous solution. Then weighing guar gum powder, adding guar gum powder into the solution with a screen while stirring, stirring at 800rpm for 4 hr to obtain 200mL of a first mixed solution, wherein Co (CH) is added₃COO)₂Is 0.11mol/L, Ca (NO)₃)₂·4H₂The concentration of O is 0.05mol/L, the concentration of disodium hydrogen phosphate is 0.03mol/L, and the concentration of guar gum is 5 g/L.

(2) The first mixed solution was injected into 200mL of 3mol/L NaOH solution at a rate of 6mL/s by a syringe to form gel fibers, and the solution was maintained for further 24 hours. Then dried at 80 ℃ for 3 hours to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 1000 ℃ at a heating rate of 15 ℃/min in the air atmosphere, preserving the heat for 3 hours, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying for 6 hours at 100 ℃, and obtaining the Co ion doped hydroxyapatite fiber material with an ultra-long structure after drying, wherein the length is about 50-60 cm, and the diameter is about 0.2 mm.

Example 7

(1) Firstly, taking BaCl₂、CaCl₂、Na₂HPO₄The solid was added to deionized water and stirred at 1000rpm for 2 hours to give a homogeneous solution. Then weighing guar gum powder, adding guar gum powder into the solution with a screen while stirring, stirring at 1500rpm for 5 hr to obtain 200mL first mixed solution containing BaCl₂The concentration of (A) is 0.064mol/L, CaCl₂The concentration of the compound is 1mol/L, the concentration of disodium hydrogen phosphate is 1mol/L, and the concentration of guar gum is 6 g/L.

(2) The first mixed solution was injected into 2000mL of 5mol/L NaOH solution at a rate of 10mL/s by a syringe to form gel fibers, and the solution was maintained for further 24 hours. Then dried at 80 ℃ for 3 hours to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 1000 ℃ at a heating rate of 15 ℃/min in the air atmosphere, preserving the heat for 3 hours, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying for 6 hours at 100 ℃, and obtaining the Ba ion doped hydroxyapatite fiber material with an ultra-long structure after drying, wherein the length is about 80-100 cm, and the diameter is about 0.2 mm.

Example 8

(1) Firstly taking FeCl₃、CaCl₂、Na₂HPO₄The solid was added to deionized water and stirred at 1000rpm for 2 hours to give a homogeneous solution. Then weighing guar gum powder, and stirring while sieving with a screenAdding into the solution, stirring at 2000rpm for 5 hr to obtain 200mL of first mixed solution containing FeCl₃The concentration of (2) is 0.1mol/L, CaCl₂The concentration of the compound is 1mol/L, the concentration of disodium hydrogen phosphate is 1mol/L, and the concentration of guar gum is 10 g/L.

(2) The first mixed solution was injected into 100mL of 5mol/L NaOH solution at a rate of 5mL/s by a syringe to form gel fibers, and the solution was maintained for further 24 hours. Then dried at 80 ℃ for 3 hours to obtain a fiber precursor.

(3) And (3) putting the fiber precursor into a crucible, putting the crucible into a high-temperature furnace, heating to 1000 ℃ at a heating rate of 15 ℃/min in the air atmosphere, preserving the heat for 3 hours, and naturally cooling to room temperature to obtain the product. Directly washing the product to be neutral by using a large amount of deionized water, then placing the washed product in an oven for drying for 6 hours at 100 ℃, and obtaining the Fe ion doped hydroxyapatite fiber material with an ultra-long structure after drying, wherein the length is about 80-90 cm, and the diameter is about 0.2 mm.

Comparative example 1

The procedure was the same as in example 1, except that guar gum powder was changed to soluble starch powder. The step (2) can not form fiber gel and can not be carried out subsequent operation.

Comparative example 2

The procedure was the same as in example 1, except that guar gum powder was changed to sodium carboxymethylcellulose powder. The step (2) can not form fiber gel and can not be carried out subsequent operation.

Second, the influence of the fibrous material on the proliferation behavior of mesenchymal stem cells

Example 9

The influence of the prepared fiber material on the proliferation behavior of the mesenchymal stem cells is measured. The measurement method is as follows:

the preparation method of the porous scaffold comprises the following steps: firstly taking Mn (NO)₃)₂、Ca(NO₃)₂·4H₂O、Na₂HPO₄The solid was added to deionized water and stirred at 200rpm for 2 hours to give a homogeneous solution. Then weighing guar gum powder, and stirringAdding guar gum powder uniformly into the solution with a screen, stirring at 200rpm for 3 hr to obtain 200mL of a first mixed solution containing Mn (NO)₃)₂Has a concentration of 0.0007mol/L, Ca (NO)₃)₂·4H₂The concentration of O is 0.5mol/L, the concentration of disodium hydrogen phosphate is 0.3mol/L, and the concentration of guar gum is 5 g/L. Injecting the first mixed solution into 100mL of 1mol/L sodium hydroxide solution at the speed of 5mL/s by using an injector to form gel fibers; after standing for 1 hour, transferring the gel fiber into a flat-bottom test tube with the diameter of 8mm, continuing standing for 2 hours, and then transferring into a refrigerator with the temperature of 20 ℃ below zero for freezing for 5 hours. Vacuum drying the frozen gel fiber for 1 day, and placing in a high temperature furnace (temperature rising speed: 5 ℃/min, 1000 ℃, heat preservation for 3 hours) to obtain the Mn-doped porous hydroxyapatite scaffold.

The preparation method of the undoped hydroxyapatite scaffold is the same as that of the undoped hydroxyapatite scaffold, but manganese nitrate is not added in the system.

The preparation method of the hydroxyapatite block scaffold comprises the following steps: directly pressing the hydroxyapatite powder synthesized by a precipitation method into a wafer with the diameter of 1cm and the thickness of 2mm by a grinding tool, and then carrying out high-temperature treatment to obtain the hydroxyapatite ceramic wafer. The high temperature treatment conditions are consistent with the conditions used in the porous scaffold preparation process.

Details of cell experiments: mouse mesenchymal stem cells are cultured in a DMEM medium containing 10% fetal calf serum, and the environment of the incubator is as follows: 5% carbon dioxide, 37 ℃ humid environment; the number of cell generations used in the experiment was 3-4, and the initial seeding density of the cells was 1 x 10⁴Per cm²Transfer of sterilized block scaffolds, Mn-doped and undoped porous scaffolds into 12-well plates, pre-soaking with culture medium for one day, followed by removal of culture medium, and cells at 5000/cm²The cell proliferation process is detected after the cell is cultured for a certain time.

As can be seen from FIG. 6, the proliferation of cells on the surface of the three scaffolds is in a gradually increasing state with the increase of the culture time, which indicates that the three scaffolds have good biocompatibility; compared with a block scaffold, the porous scaffold has a more obvious effect of promoting cell proliferation; when Mn is introduced into the scaffold, the cells have higher proliferation speed on the surface of the Mn-doped porous scaffold and show higher light absorption values, which indicates that Mn doping can further promote the proliferation of the cells on the surface of the scaffold.

The metal ion doped/undoped scaffolds (gel fibers were transferred to a flat bottom tube with a diameter of 8mm before drying) were prepared according to the parameters of examples 3 and 4, respectively, and similar results as described above were obtained, and the promotion effect of the porous scaffold on cell proliferation behavior was more significant, and the data are not listed.

The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. When other parameters defined by the present invention are adopted, the technical effects disclosed in the present specification can be obtained, and the details are not repeated herein for the sake of brevity. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Claims

1. A preparation method of a metal ion doped hydroxyapatite fiber material comprises the following steps:

(1) mixing ion-doped salt, soluble calcium salt, guar gum and disodium hydrogen phosphate with deionized water to obtain a first mixed solution, wherein the concentration of the ion-doped salt is more than 0 and less than or equal to 0.1 mol/L;

(3) and (3) calcining the fiber precursor obtained in the step (2) to obtain a product, washing the product with deionized water, and drying to obtain the metal ion-doped hydroxyapatite fiber material.

2. The method of claim 1, wherein in step (1), the salt of the dopant ion is a salt of magnesium, strontium, barium, manganese, cobalt, iron, nickel, copper, or zinc selected from chloride, nitrate, or acetate; the soluble calcium salt is calcium nitrate or calcium chloride.

3. The method according to claim 1 or 2, wherein the first mixed solution obtained in the step (1) has a soluble calcium salt concentration of 0.005 to 1mol/L, a disodium hydrogen phosphate concentration of 0.005 to 1mol/L, and a guar gum concentration of 0.1 to 10 g/L.

4. The method of claim 1 or 2, wherein in the step (2), the strong alkali solution is a sodium hydroxide solution or a potassium hydroxide solution, and the concentration is 0.1-10 mol/L; the injection speed is 1-10 mL/s; the soaking time is 1-240 hours; the volume ratio of the first mixed solution to the strong base solution is 1: 10-10: 1.

5. The method of claim 1 or 2, wherein in step (2), the drying process conditions are: drying for 2-72 hours at 40-120 ℃; or vacuum drying for 1-2 days at-80 to-20 ℃.

6. The method of claim 1 or 2, wherein in step (3), the calcination process conditions are: the heating rate is 1-20 ℃/min, the temperature is increased from room temperature to 800-1250 ℃, the temperature is preserved for 1-6 hours, and then the product is naturally cooled to room temperature.

7. The method of claim 1 or 2, wherein in step (3), the drying process conditions are: the drying temperature is room temperature to 300 ℃, and the drying time is 1 to 72 hours.

8. A metal ion doped hydroxyapatite fibrous material prepared according to the method of any one of claims 1 to 7.

9. A hydroxyapatite scaffold, characterized by consisting of the metal ion doped hydroxyapatite fiber material according to claim 8.