Refined manufacturing of photo-control-hybridized cross-linked degradable support and bone tissue engineering application of photo-control-hybridized cross-linked degradable support

Abstract

The invention belongs to the technical field of biomaterials and particularly relates to refined manufacturing of a photo-control-hybridized cross-linked degradable support and bone tissue engineeringapplication of the photo-control-hybridized cross-linked degradable support. A method for preparing the photo-control-hybridized cross-linked degradable support comprises the steps: 1) preparing modified hydroxyapatite; 2) preparing a bonder GelMA; and 3) blending a blue-light initiator and a thickener to prepare slurry, subjecting double bonds in the GelMA and double bonds of silane-coupler-modified grafted hydroxyapatite to an interaction, i.e., a formula shown in the description in the presence of free radicals through 3D printing technique forming under the condition of illumination of specific wavelength, then, initiating monomer polymerization, and carrying out bonding, thereby forming the solid-phase hybridized degradable support. The invention relates to application of the osteogenesis-remediation-promoting photo-crosslinked composite 3D-printed hybridized degradable support as a drug carrier or / and a bioscaffold material. The support is scientific in design and simple and convenient in operation and can be applied to carrier support materials of bone-defected regenerated remediation or all classes of bioactive substances due to refined controllable processing characteristics and biodegradable characteristics.

Description

technical field

[0001] The invention belongs to the technical field of biomaterials, in particular to the refined manufacture of light-controlled hybrid cross-linked degradable scaffolds and its application in bone tissue engineering. Background technique

[0002] Tissue engineering is an important emerging topic in the field of biomedical engineering, and its three elements are seed cells, scaffolds and growth factors. As one of the three elements of tissue engineering, scaffold plays an important role in it. Generally speaking, both the scaffold itself and its degradation products should have good biocompatibility. Moreover, the scaffold needs to have suitable mechanical properties to provide a good stress environment for the new tissue. In addition, the scaffold should possess a perforated porous structure and permeability to allow the ingress of cells and nutrients and the egress of metabolic wastes. Finally, the scaffold needs to have suitable surface structure and p...

Examples

Embodiment 1

[0088] This embodiment discloses a method for preparing a double bond modified hydroxyapatite slurry, and the specific steps are as follows:

[0089] (1) Dosing: the volume ratio of ethanol and water is 6:1, 7:1, 8:1, and 9:1 respectively to obtain a solution, and the pH value is adjusted to 4.0 with glacial acetic acid;

[0090] In this step, the ethanol:water volume ratios are 6:1, 7:1, 8:1, and 9:1, which are recorded as experimental groups ①, ②, ③, and ④ respectively.

[0091] (2) Hydrolysis reaction: Take the ethanol-water solution prepared in step (1), stir it with a magnetic stirrer, and slowly add 5ml of ethyl orthosilicate and vinyltriethoxysilane in a volume ratio of 1:1. solution, stirred at room temperature for 1 h;

[0092] (3) Condensation reaction: slowly drop 20ml of hydroxyapatite slurry into the mixed solution prepared in step (2), stir and react at room temperature for 1.5h to obtain a mixed solution;

[0093] (4) Add dropwise 10wt% NaOH solution to the mi...

Embodiment 2

[0104] This embodiment discloses the preparation method of methacrylic anhydride gelatin, and concrete steps are as follows:

[0105] (1) Dosing: Dissolve gelatin in PBS buffer at a concentration of 10% (w / v), and stir well at 50°C until the gelatin is completely dissolved;

[0106] (2) Add methacrylic anhydride MA (0.8ml~1.2mLMA per gram of gelatin) dropwise to the gelatin solution prepared in step (1) at a rate of 0.5ml per minute, and fully stir the reaction while maintaining 50°C 3h;

[0107] In this step, the ratio (v / w) of methacrylic anhydride to gelatin is: 0.6:1, 0.8:1, 1:1, 1.2:1. Record them as experimental groups A, B, C, and D respectively;

[0108] (3) After the reaction is completed, the product obtained in step 2 is transferred to a centrifuge tube, centrifuged at 1500rpm for 5min, and the supernatant is taken and packed into a dialysis bag;

[0109] (4) Dialysis: dialyze the product obtained in step 3 in pure water at 50° C. for at least 3 days, and change ...

Embodiment 3

[0117] The GELMA of experimental group C was configured as a hydrogel with a concentration of 15% (w / v) and placed on a dynamic mechanical analyzer (TAInstruments Q800, USA), and the hydrogel was measured in the multi-frequency mode (0.1-100rad / s). The change curves of elastic modulus (G′) and viscous modulus (G″), the results are shown in the attached Figure 6 shown. Under the test condition that the amplitude is fixed at 0.5%, the frequency is in the range of 0.1-100rad / s, the elastic modulus G' is greater than the viscous modulus G", the material shows obvious hydrogel characteristics, and the elastic modulus G ′ and viscosity modulus G″ both show a strong frequency dependence, which increases with the increase of frequency, and the values ​​are 0.01Pa~139Pa and 0.01Pa~72Pa, respectively.

technical field

[0001] The invention belongs to the technical field of biomaterials, in particular to the refined manufacture of light-controlled hybrid cross-linked degradable scaffolds and its application in bone tissue engineering. Background technique

[0002] Tissue engineering is an important emerging topic in the field of biomedical engineering, and its three elements are seed cells, scaffolds and growth factors. As one of the three elements of tissue engineering, scaffold plays an important role in it. Generally speaking, both the scaffold itself and its degradation products should have good biocompatibility. Moreover, the scaffold needs to have suitable mechanical properties to provide a good stress environment for the new tissue. In addition, the scaffold should possess a perforated porous structure and permeability to allow the ingress of cells and nutrients and the egress of metabolic wastes. Finally, the scaffold needs to have suitable surface structure and p...

Examples

Embodiment 1

[0088] This embodiment discloses a method for preparing a double bond modified hydroxyapatite slurry, and the specific steps are as follows:

[0089] (1) Dosing: the volume ratio of ethanol and water is 6:1, 7:1, 8:1, and 9:1 respectively to obtain a solution, and the pH value is adjusted to 4.0 with glacial acetic acid;

[0090] In this step, the ethanol:water volume ratios are 6:1, 7:1, 8:1, and 9:1, which are recorded as experimental groups ①, ②, ③, and ④ respectively.

[0091] (2) Hydrolysis reaction: Take the ethanol-water solution prepared in step (1), stir it with a magnetic stirrer, and slowly add 5ml of ethyl orthosilicate and vinyltriethoxysilane in a volume ratio of 1:1. solution, stirred at room temperature for 1 h;

[0092] (3) Condensation reaction: slowly drop 20ml of hydroxyapatite slurry into the mixed solution prepared in step (2), stir and react at room temperature for 1.5h to obtain a mixed solution;

[0093] (4) Add dropwise 10wt% NaOH solution to the mi...

Embodiment 2

[0104] This embodiment discloses the preparation method of methacrylic anhydride gelatin, and concrete steps are as follows:

[0105] (1) Dosing: Dissolve gelatin in PBS buffer at a concentration of 10% (w / v), and stir well at 50°C until the gelatin is completely dissolved;

[0106] (2) Add methacrylic anhydride MA (0.8ml~1.2mLMA per gram of gelatin) dropwise to the gelatin solution prepared in step (1) at a rate of 0.5ml per minute, and fully stir the reaction while maintaining 50°C 3h;

[0107] In this step, the ratio (v / w) of methacrylic anhydride to gelatin is: 0.6:1, 0.8:1, 1:1, 1.2:1. Record them as experimental groups A, B, C, and D respectively;

[0108] (3) After the reaction is completed, the product obtained in step 2 is transferred to a centrifuge tube, centrifuged at 1500rpm for 5min, and the supernatant is taken and packed into a dialysis bag;

[0109] (4) Dialysis: dialyze the product obtained in step 3 in pure water at 50° C. for at least 3 days, and change ...

Embodiment 3

[0117] The GELMA of experimental group C was configured as a hydrogel with a concentration of 15% (w / v) and placed on a dynamic mechanical analyzer (TAInstruments Q800, USA), and the hydrogel was measured in the multi-frequency mode (0.1-100rad / s). The change curves of elastic modulus (G′) and viscous modulus (G″), the results are shown in the attached Figure 6 shown. Under the test condition that the amplitude is fixed at 0.5%, the frequency is in the range of 0.1-100rad / s, the elastic modulus G' is greater than the viscous modulus G", the material shows obvious hydrogel characteristics, and the elastic modulus G ′ and viscosity modulus G″ both show a strong frequency dependence, which increases with the increase of frequency, and the values ​​are 0.01Pa~139Pa and 0.01Pa~72Pa, respectively.

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