CN111803706A - Preparation method of bone-like bioactive polycaprolactone porous scaffold and porous scaffold - Google Patents
Preparation method of bone-like bioactive polycaprolactone porous scaffold and porous scaffold Download PDFInfo
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Abstract
The invention discloses a preparation method of a bone-imitated bioactive polycaprolactone porous scaffold and a porous scaffold, which comprises the following steps of (1) preparing the polycaprolactone porous scaffold; (2) construction of the patterned surface: immersing the porous scaffold prepared in the step (1) into a growth liquid (in which a proper amount of polycaprolactone is dissolved), taking out, naturally volatilizing the solvent, and forming a uniformly distributed patterned structure on the surface and inside of the porous scaffold through epiphytic crystals; (3) generation of a bone-like bioactive layer: and (3) immersing the porous support with the patterned structure prepared in the step (2) into a calcium-phosphorus precursor solution, adjusting the pH value of the calcium-phosphorus precursor solution to be alkalescent by using a pH regulator, inducing and accelerating the deposition generation of the bone-like hydroxyapatite coating by using the high specific surface area of the patterned structure, then cleaning the amorphous sediment on the surface of the porous support, and drying in vacuum to obtain the bone-like bioactive porous support. The scaffold constructed by the invention has high deposition efficiency of the bone-like calcium-phosphorus layer and good biocompatibility.
Description
Technical Field
The invention relates to the field of preparation of high-molecular bone tissue engineering scaffolds, in particular to a preparation method of an imitated bone bioactive polycaprolactone porous scaffold and the porous scaffold.
Background
Bone damage or bone defects caused by accidents or diseases become one of the important problems in orthopedics, and the number of bone grafting operations is over 200 ten thousand every year worldwide, and the treatment amount is nearly $ 25 hundred million. Up to now, autologous bone grafting is the "gold standard" for treating bone defects, however, autologous bone is not available, secondary damage is easily caused, and complications (such as mucosal dehiscence, wound dehiscence and the like) are caused. While the foreign body bone is easy to cause infection, the application of the bone is also limited. With advances in the medical level, Bone Tissue Engineering (BTE) technology provides a new approach to the treatment of bone injuries. The bone tissue engineering scaffold is the key of BTE, and the ideal bone tissue engineering scaffold has good biocompatibility, high porosity and proper pore size to meet the requirements of nutrient exchange of cells, in vivo degradation, excellent osteoconductivity and the like. Biodegradable medical polymer materials, such as polylactic acid, Polycaprolactone (PCL), etc., have been widely used as scaffold materials for bone tissue engineering due to their good biocompatibility, excellent processability, good mechanical matching properties, etc. However, the biological inertness of the polymer material itself also limits the in vivo repair effect of the porous scaffold.
Hydroxyapatite (HA) is the main inorganic component of natural bone and HAs good biocompatibility and bone conduction performance. In order to improve the interface problem between cells and polymer scaffolds, the biomedical engineering field HAs been dedicated to introducing HA layers on the surfaces of polymer materials for many years. Generally, the HA layer is formed by combining calcium and phosphorus ions in the precursor solution into amorphous calcium and phosphorus composite salt under a weak alkaline condition, and combining the amorphous calcium and phosphorus composite salt with nucleation sites on the surface of the stent to further form the HA layer. The HA layer provides an effective binding surface to enhance the bioactivity of the material and promote osteogenic differentiation of cells [ M Bosetti et al, Biomaterials2005, 26, 7572 ]. The Chinese patent of invention (publication No. CN 110302424A) coats dopamine coating on the surface of metal or macromolecule support, which can firmly adhere to the substrate, and then mineralizes in supersaturated calcium phosphate solution array containing to-be-doped element ions for 1-10 days to obtain multi-element doped calcium phosphate coating. The Chinese patent of invention (publication No. CN 109675104A) irradiates photoactivated biomacromolecules and photoactivated phosphatase with ultraviolet and visible light with the wavelength of 320-500 nm to form hydrogel in a crosslinking way, and the phosphatase can promote phosphate deposition to obtain the uniformly calcified hydrogel. The Chinese invention patent (publication No. CN 107224609A) finds that the zwitterionic polyelectrolyte has a stabilizing effect on calcium phosphate solution. The degradable bionic calcified collagen scaffold which is similar to a natural bone tissue structure, has good mechanical property and biocompatibility is prepared by forming calcification through collagen fibers loaded with amphoteric polyelectrolyte.
Most of the high molecular materials are hydrophobic and biologically inert, and calcium ions and phosphate ions are difficult to nucleate and calcify on the surfaces of the high molecular materials. At present, hydrophilic functional groups can be introduced on the surface of a high polymer material through complex chemical reactions to promote nucleation, but the chemical modification operation is complex, additional chemical substances are easily introduced in the process, and side effects on human bodies still need to be observed. In contrast, the surface physical modification method is a simple and effective method for promoting calcium phosphorus deposition. According to the method, a micro-nano structure is usually constructed on the surface of a high polymer material, so that the surface roughness is improved, and the activation energy of deposition nucleation is reduced. For example, increasing the surface roughness of polyetheretherketone by sandblasting can increase the amount of surface deposition and improve the material bioactivity [ H Mahjoubi et al Acta biomaterials, 2017, 47,149 ]. Zhang et al regulating polypeptides are self-assembled to obtain films with different surface structures, and the discovery shows that the rough porous surface can load more HA nanoparticles, which is beneficial to osteogenic differentiation of stem cells [ Zhang et al Nanoscale 2017, 9, 13670 ].
At present, methods for constructing surface micro-nano structures also include template methods, laser etching and the like, but the methods are only suitable for two-dimensional planes. In tissue engineering, the three-dimensional porous scaffold simulates extracellular matrix, provides temporary growth environment for cells and is a necessary condition for cell growth.
To this end, it is desirable to seek a solution that at least alleviates the above problems.
Disclosure of Invention
The invention aims to provide a preparation method of a bone-like bioactive polycaprolactone porous scaffold, which can be used for constructing uniform micro-nano structures on the surface and in the porous scaffold, promoting the construction of a bone-like bioactive layer and preparing a high-molecular porous scaffold with good bioactivity.
The technical scheme of the invention for realizing the purpose is as follows:
a preparation method of a bone-imitated bioactive polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: preparing a porous scaffold from polycaprolactone;
(2) construction of the patterned surface: immersing the porous scaffold prepared in the step (1) into a growth liquid (in which a proper amount of polycaprolactone is dissolved), taking out, naturally volatilizing the solvent, and forming a uniformly distributed patterned structure on the surface and inside of the porous scaffold through epiphytic crystals;
(3) generation of a bone-like bioactive layer: and (3) immersing the porous support with the patterned structure prepared in the step (2) into a calcium-phosphorus precursor solution, adjusting the pH value of the calcium-phosphorus precursor solution to be alkalescent by using a pH regulator, inducing and accelerating the deposition generation of the bone-like hydroxyapatite coating by using the high specific surface area of the patterned structure, then cleaning the amorphous sediment on the surface of the porous support taken out of the calcium-phosphorus precursor solution, and drying in vacuum to obtain the bone-like bioactive porous support.
The step (1) is specifically as follows: and extruding the dried polycaprolactone granules to obtain printing wires with uniform diameters, and then preparing the porous scaffold by using a fused deposition 3D printing technology.
The polycaprolactone is biomedical grade and has the molecular weight of 60000 g/mol.
In step (1), the size that 3D printed porous support hole is 500 +/-50 um, and the printing line diameter is 300 +/-30 um.
The molar ratio of the components of the calcium-phosphorus precursor solution is as follows: NaCl, KCl, CaCl2ּ·2H2O: MgCl2:NaH2PO4=200:1:5:1:2。
The concentration of the calcium-phosphorus precursor solution is 1-10 times of that of a standard SBF solution.
In the step (2), the growth liquid is a solution formed by dissolving polycaprolactone in an acetic acid/water mixed solution.
The volume ratio of the acetic acid to the water is 3: 1.
In the growth solution, the solubility of polycaprolactone in the acetic acid/water mixed solution is 0.5-5% w/v.
In the step (2), the time of the epiphytic crystallization is 5-60 min.
In the step (3), the PH regulator is 0.1mol/L tris-HCl buffer solution; the pH of the calcium-phosphorus precursor solution is adjusted to be 7.4 +/-0.1.
In the step (3), the deposition time is 5-48 h.
The beneficial technical effects of the invention are as follows:
according to the invention, the surface patterning structure is constructed in a physical mode, so that the surface roughness of the stent material is improved, and nucleation sites are provided for combining calcium and phosphorus ions. The invention utilizes solution homogeneous phase epitaxial crystallization to generate upright lamella (Edge-on lamellae) on the surface and inside the macromolecular porous bracket, and effectively promotes the formation of a uniform and compact bone-like bioactive layer. The method is simple and efficient, is suitable for the complex surface and the interior of the three-dimensional porous scaffold, and has good universality. In addition, the bone-like bioactive polycaprolactone porous scaffold prepared by the method has good bioactivity, can remarkably promote the proliferation and osteogenic differentiation of mouse bone marrow mesenchymal stem cells, and has good application potential in the aspect of bone tissue engineering.
Compared with the prior art, the method has the following advantages:
1. the invention provides a method for effectively promoting the generation of a calcium-phosphorus bioactive layer, which is simple and efficient to operate, does not introduce other components, and ensures the biocompatibility of the stent material.
2. The patterned surface capable of promoting calcification consists of upright platelets generated by homogeneous periphytic crystallization, and strong interaction is generated between the patterned surface and the substrate through lattice matching, so that interface bonding is improved, and the interface stability of the calcium-phosphorus bioactive layer is also ensured.
3. The invention can realize the construction of the bone-like bioactive layer on the surface and in the three-dimensional porous bracket and has good universality.
Drawings
FIG. 1 is a schematic diagram of the preparation process of the bone-like bioactive polycaprolactone porous scaffold.
FIG. 2 is a scanning electron microscope image of the surface of the porous scaffolds prepared in comparative example 1, comparative example 2, comparative example 3 and example 10 according to the present invention.
Fig. 3 is a alizarin red staining diagram of the porous scaffolds prepared in comparative example 3 and example 6 of the present invention.
Fig. 4 is a graph showing the thermal weight loss of the surfaces of the porous scaffolds prepared in comparative example 1, comparative example 2, comparative example 3 and example 10 according to the present invention.
FIG. 5 is a graph showing the results of quantifying CCK-8 in the cells cultured on the porous scaffolds prepared in comparative example 1, comparative example 2, comparative example 3 and example 10.
FIG. 6 is a graph showing the results of qRT-PCR after 7 days of culturing the cells in the porous scaffolds prepared in comparative example 1, comparative example 2, comparative example 3 and example 10 according to the present invention.
Detailed Description
The following examples are given to illustrate the present invention and it is necessary to point out here that the following examples are given only for the purpose of further illustration of the invention and are not to be construed as limiting the scope of the invention.
Table 1 preparation process conditions and deposition amounts of examples 1 to 12 and comparative examples 1 to 3.
Examples 1 to 4
Referring to table 1, the preparation method of the bone-like bioactive polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous support by using a fused deposition 3D printing technology;
(2) construction of the patterned surface: respectively immersing the porous scaffold prepared in the step (1) into growth liquid dissolved with 0.5, 1.0, 2.0 and 5.0% w/v of PCL for 5min, taking out, naturally volatilizing the solvent, and generating uniform patterned structures on the surface and inside of the scaffold through epiphytic crystallization;
(3) generation of a bone-like bioactive layer: and (3) immersing the porous support with the patterned structure prepared in the step (2) into 1 x calcium phosphorus precursor liquid, adjusting the pH value of the calcium phosphorus precursor liquid to alkalescence through a pH regulator, inducing and accelerating the deposition generation of the bionic hydroxyapatite coating by utilizing the high specific surface area of the patterned structure, depositing for 5 hours, then cleaning the amorphous sediment on the surface of the porous support taken out of the calcium phosphorus precursor liquid, and drying in vacuum to obtain the bone-like bioactive porous support.
Examples 5 to 7
Referring to table 1, the preparation method of the bone-like bioactive polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous support by using a fused deposition 3D printing technology;
(2) construction of the patterned surface: respectively immersing the hole support prepared in the step (1) into growth liquid dissolved with 0.5wt% of PCL for 10min, 30 min and 60min, taking out, naturally volatilizing the solvent, and generating uniform patterned structures on the surface and inside of the support through epiphytic crystallization;
(3) generation of a bone-like bioactive layer: and (3) immersing the porous support with the patterned surface prepared in the step (2) into 1 x calcium phosphorus precursor solution, adjusting the pH value of the calcium phosphorus precursor solution to be alkalescent by using a pH regulator, inducing and accelerating the deposition generation of the bionic hydroxyapatite coating by using the high specific surface of the patterned surface, depositing for 5 hours, then cleaning the amorphous sediment on the surface of the porous support taken out of the calcium phosphorus precursor solution, and drying in vacuum to obtain the bone-like bioactive porous support.
Examples 8 to 9
Referring to table 1, the preparation method of the bone-like bioactive polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous support by using a fused deposition 3D printing technology;
(2) construction of the patterned surface: immersing the porous scaffold prepared in the step (1) in a growth solution of 0.5wt% of PCL for 5min, taking out, volatilizing the solvent under natural conditions, and generating uniform patterned structures on the surface and inside of the scaffold through epiphytic crystallization;
(3) generation of a bone-like bioactive layer: and (3) respectively immersing the porous scaffold with the patterned surface prepared in the step (2) into 5 x calcium phosphorus precursor liquid and 10 x calcium phosphorus precursor liquid, adjusting the pH value of the calcium phosphorus precursor liquid to be alkalescent through a pH regulator, inducing and accelerating the deposition generation of the bionic hydroxyapatite coating by utilizing the high specific surface area of the patterned structure, depositing for 5 hours, then cleaning the amorphous sediment on the surface of the porous scaffold taken out of the calcium phosphorus precursor liquid, and drying in vacuum to obtain the bone-like bioactive porous scaffold.
Examples 10 to 12
Referring to table 1, the preparation method of the bone-like bioactive polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous PCL support by using a fused deposition 3D printing technology;
(2) construction of the patterned surface: immersing the porous scaffold prepared in the step (1) in a growth solution of 0.5wt% of PCL for 5min, taking out, naturally volatilizing the solvent, and enabling the surface and the interior of the scaffold to generate a uniform patterned structure through epiphytic crystallization;
(3) generation of a bone-like bioactive layer: and (3) immersing the porous support with the patterned surface prepared in the step (2) into 1 x calcium phosphorus precursor solution, adjusting the pH value of the calcium phosphorus precursor solution to be alkalescent through a pH regulator, inducing and accelerating the deposition generation of the bionic hydroxyapatite coating through nano vertical lamella, respectively depositing for 12h, 24 h and 48h, then cleaning the amorphous sediment on the surface of the porous support taken out of the calcium phosphorus precursor solution, and drying in vacuum to obtain the bone-like bioactive porous support.
In the preceding examples, the polycaprolactone was biomedical grade, having a molecular weight of 60000 g/mol.
In the step (1) of the previous embodiment, the size of the pores of the 3D printing porous support is 500 +/-50 um, and the diameter of the printing line is 300 +/-30 um.
In the previous embodiment, the molar ratio of the components of the calcium-phosphorus precursor solution is as follows: NaCl, KCl, CaCl2ּ·2H2O :MgCl2:NaH2PO4=200:1:5:1:2。
In the step (2) of the previous embodiment, the growth liquid is a solution formed by dissolving polycaprolactone in a mixed solution of acetic acid and water. The ratio of acetic acid to water was 3: 1.
In step (3) of the previous example, the pH adjusting agent was 0.1mol/L tris-HCl buffer solution; the pH of the calcium-phosphorus precursor solution is adjusted to be 7.4 +/-0.1.
Comparative example 1
Referring to table 1, a preparation method of a polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: and extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous scaffold by using a fused deposition 3D printing technology as a blank control.
Comparative example 2
Referring to table 1, a preparation method of a polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous support by using a fused deposition 3D printing technology;
(2) construction of the patterned surface: and (2) immersing the porous scaffold prepared in the step (1) in a growth solution of 2wt% of PCL for 10min, taking out, naturally volatilizing the solvent, and generating uniform patterned surface structures on the surface and inside of the scaffold through epiphytic crystallization to prepare the scaffold with only the patterned surface.
Comparative example 3
Referring to table 1, a preparation method of a polycaprolactone porous scaffold comprises the following steps:
(1) preparing a porous scaffold: extruding the dried Polycaprolactone (PCL) granules to prepare printing wires with uniform diameters, and then preparing the porous PCL support by using a fused deposition 3D printing technology;
(2) generation of a bone-like bioactive layer: and (2) immersing the porous scaffold prepared in the step (1) in a precursor solution of 10 gamma, adjusting the pH value to be alkalescent, depositing for 12h, cleaning amorphous deposits on the surface of the scaffold, and drying in vacuum to obtain the porous scaffold only with the calcium-phosphorus layer.
The invention also provides a porous scaffold prepared by the preparation method of the bone-like bioactive polycaprolactone porous scaffold, and the related steps and flows are shown in figure 1.
The surface morphology of the porous scaffolds prepared in comparative examples 1, 2, and 3 and example 10 was observed using a field emission scanning electron microscope (FE-SEM), as shown in fig. 2. The untreated porous scaffold prepared in comparative example 1 had a smooth and flat surface. The porous scaffold prepared in comparative example 2 exhibited a wide range of patterned surfaces. As can be seen from the enlarged view, the patterned surface is composed of uniformly distributed nano-standing platelets, demonstrating the efficient ability of solution homoepitaxial crystallization to build up a patterned surface. The surface of the porous scaffold prepared in comparative example 3 had only a few HA deposits due to the lack of calcification nucleation sites in the smooth scaffold. In contrast, the surface of the porous scaffold prepared in example 10 formed a uniform and dense HA layer, indicating that the patterned surface built up of upstanding platelets significantly promoted the deposition of the calcium-phosphorus active layer.
The distribution of HA on the surface of the porous scaffold was observed by alizarin red staining, as shown in FIG. 3. The porous scaffold prepared in comparative example 3 had weak staining locally, while the porous scaffold prepared in example 10 was stained in its entirety and darker in color, indicating that the patterned surface composed of nano-sized upright platelets effectively promotes calcium phosphorus deposition, and the HA layer was dense and uniformly distributed, covering the entire scaffold. Fig. 4 is a graph of the thermal weight loss of the surfaces of the porous scaffolds prepared in comparative example 1, comparative example 2, comparative example 3, and example 10, and the final residual amount of the porous scaffolds prepared in comparative example 1 and comparative example 2 was nearly 0, indicating that the PCL matrix had completely decomposed after high-temperature ablation. In contrast, the porous scaffolds prepared in comparative example 3 and example 10 had some mass remaining after high temperature ablation, indicating that they were derived from the HA deposit. In particular, the HA deposition layer of the porous scaffold prepared in example 10 reaches 5.4%, which is improved by 1.25 times compared with the porous scaffold prepared in comparative example 3, and it is confirmed that the patterned surface formed by the upright platelets can effectively promote the calcium-phosphorus ion binding deposition, and the HA adhesion amount is significantly increased.
To evaluate the effect of the osteoid active layer on cell proliferation, the proliferation of bone marrow mesenchymal stem cells (BMSCs) cultured on a porous scaffold for many days was tested using a cell counting kit (CCK-8), as shown in fig. 5. The absorbance of the porous scaffold prepared in example 10 was the highest regardless of the number of days of culture, i.e., the osteoid bioactive layer was more advantageous to promote the proliferation of BMSCs. Meanwhile, in order to evaluate the capacity of the osteoactive-like porous scaffold to promote bone differentiation, the expression of osteogenesis-related proteins of BMSC was tested after 7 days of culture using quantitative reverse transcription real-time polymerase chain reaction (qRT-PCR), as shown in fig. 6. Compared with comparative example 1, the expressions of osteogenic related gene proteins such as alkaline phosphatase (ALP), Osteocalcin (OCN) and the like in comparative example 2, comparative example 3 and example 10 are all remarkably up-regulated, which indicates that the surface patterning and the construction of the active layer can promote osteogenic differentiation of BMSCs. In particular, the expression levels of the related proteins in example 10 were all at the highest level, indicating that the patterned bone-like active layer has a stronger ability to promote the differentiation of BMSCs into osteoblasts.
Therefore, the method induces and generates the vertical platelet on the 3D printing porous support through the solution homogeneous phase epitaxial crystallization, constructs the patterned surface, and improves the specific surface roughness, thereby providing sites for the adsorption and combination of calcium and phosphorus particles and effectively promoting the formation of the bone-like bioactive layer. The bone-imitating bioactive polycaprolactone porous scaffold prepared by the invention has good bioactivity, can effectively promote cell adhesion growth and osteogenic differentiation, and has good application potential in the field of bone tissue engineering scaffolds.
It should be noted that the various features described in the foregoing embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in any further detail in order to avoid unnecessary repetition.
The present invention has been described in detail with reference to the embodiments, which are illustrative rather than restrictive, and variations and modifications thereof are possible within the scope of the present invention without departing from the general inventive concept.
Claims (13)
1. A preparation method of a bone-like bioactive polycaprolactone porous scaffold is characterized by comprising the following steps:
(1) preparing a porous scaffold: preparing a porous scaffold from polycaprolactone;
(2) construction of the patterned surface: immersing the porous scaffold prepared in the step (1) into a growth solution dissolved with a proper amount of polycaprolactone, taking out the porous scaffold, naturally volatilizing the solvent, and forming a uniform patterned structure on the surface and inside the porous scaffold through epiphytic crystallization;
(3) generation of a bone-like bioactive layer: and (3) immersing the porous support with the patterned structure prepared in the step (2) into a calcium-phosphorus precursor solution, adjusting the pH value of the calcium-phosphorus precursor solution to be alkalescent by using a pH regulator, inducing and accelerating the deposition generation of the bone-like hydroxyapatite coating by using the high specific surface area of the patterned structure, then cleaning the amorphous sediment on the surface of the porous support taken out of the calcium-phosphorus precursor solution, and drying in vacuum to obtain the bone-like bioactive porous support.
2. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein the step (1) is specifically as follows: and extruding the dried polycaprolactone granules to obtain printing wires with uniform diameters, and then preparing the porous scaffold by using a fused deposition 3D printing technology.
3. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein the polycaprolactone is biomedical grade and has a molecular weight of 60000 g/mol.
4. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 2, wherein in the step (1), the size of the pores of the 3D-printed porous scaffold is 500 +/-50 um, and the diameter of the printed lines is 300 +/-30 um.
5. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein the molar ratio of the components of the calcium-phosphorus precursor solution is as follows: NaCl, KCl, CaCl2ּ·2H2O:MgCl2:NaH2PO4=200:1:5:1:2。
6. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein the concentration of the calcium-phosphorus precursor solution is 1-10 times that of a standard SBF solution.
7. The method for preparing the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein in the step (2), the growth liquid is a solution formed by dissolving polycaprolactone in an acetic acid/water mixed solution.
8. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 7, wherein the volume ratio of the acetic acid to the water is 3: 1.
9. The method for preparing the porous scaffold of the biomimetic bone bioactive polycaprolactone according to claim 7, wherein the concentration of polycaprolactone in the acetic acid/water mixed solution in the growth solution is 0.5-5% w/v.
10. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein in the step (2), the time for epitopic crystallization is 5-60 min.
11. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein in the step (3), the pH regulator is 0.1mol/L tris-HCl buffer solution, and the pH of the calcium-phosphorus precursor solution is adjusted to 7.4 +/-0.1.
12. The preparation method of the bone-like bioactive polycaprolactone porous scaffold according to claim 1, wherein in the step (3), the deposition time is 5-48 h.
13. A porous scaffold, which is prepared by the preparation method of the bone-like bioactive polycaprolactone porous scaffold of any one of claims 1-12.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113730658A (en) * | 2021-09-22 | 2021-12-03 | 太原理工大学 | Bionic bone cartilage integrated repair support and preparation method thereof |
| CN117565386A (en) * | 2024-01-17 | 2024-02-20 | 中国科学院化学研究所 | Cell or organoid chip and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104623728A (en) * | 2015-01-12 | 2015-05-20 | 四川大学 | Injection molding anisotropic bone substitute composite material and preparation method thereof |
| CN106474566A (en) * | 2016-10-19 | 2017-03-08 | 深圳市艾科赛龙科技股份有限公司 | A kind of 3D printing PCL/HAp composite and preparation method thereof, purposes, Method of printing |
| CN106620879A (en) * | 2015-10-29 | 2017-05-10 | 张智勇 | Bionic bone repair material having good machinery performance and biological activity and preparation method thereof |
| WO2019018443A1 (en) * | 2017-07-17 | 2019-01-24 | Stc.Unm | Scaffolds for bone-soft tissue interface and methods of fabricating the same |
-
2020
- 2020-07-04 CN CN202010633177.0A patent/CN111803706A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104623728A (en) * | 2015-01-12 | 2015-05-20 | 四川大学 | Injection molding anisotropic bone substitute composite material and preparation method thereof |
| CN106620879A (en) * | 2015-10-29 | 2017-05-10 | 张智勇 | Bionic bone repair material having good machinery performance and biological activity and preparation method thereof |
| CN106474566A (en) * | 2016-10-19 | 2017-03-08 | 深圳市艾科赛龙科技股份有限公司 | A kind of 3D printing PCL/HAp composite and preparation method thereof, purposes, Method of printing |
| WO2019018443A1 (en) * | 2017-07-17 | 2019-01-24 | Stc.Unm | Scaffolds for bone-soft tissue interface and methods of fabricating the same |
Non-Patent Citations (4)
| Title |
|---|
| HUA-MO YIN等: "Surface Epitaxial Crystallization-Directed Nanotopography for Accelerating Preosteoblast Proliferation and Osteogenic Differentiation", 《ACS APPL. MATER. INTERFACES》 * |
| SU A PARK等: "Fabrication of Biomimetic PCL Scaffold Using Rapid Prototyping for Bone Tissue Engineering", 《MACROMOLECULAR RESEARCH》 * |
| 叶福丽等: "钛表面不同凝胶层对磷酸钙生物活性层形成的影响 ", 《咸宁学院学报》 * |
| 王迎军主编: "《新型材料科学与技术 无机材料卷 下》", 31 October 2016, 华南理工大学出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113730658A (en) * | 2021-09-22 | 2021-12-03 | 太原理工大学 | Bionic bone cartilage integrated repair support and preparation method thereof |
| CN113730658B (en) * | 2021-09-22 | 2022-08-16 | 太原理工大学 | Bionic bone cartilage integrated repair support and preparation method thereof |
| CN117565386A (en) * | 2024-01-17 | 2024-02-20 | 中国科学院化学研究所 | Cell or organoid chip and preparation method and application thereof |
| CN117565386B (en) * | 2024-01-17 | 2024-03-22 | 中国科学院化学研究所 | Cell or organoid chip and preparation method and application thereof |
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