CN114854076A - Bioactive SBS material for skull restoration and preparation method and application thereof - Google Patents
Bioactive SBS material for skull restoration and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a bioactive SBS material for skull restoration and a preparation method and application thereof. The preparation method comprises the following steps: covalently connecting the modified quaternary ammonium salt chitosan on an SBS substrate, and then fixing a polydopamine modified zeolite imidazole ester skeleton-8 and polydopamine modified hydroxyapatite on the surface of the modified quaternary ammonium salt chitosan. The skull forming material prepared by the invention has the inherent bactericidal performance and the capability of promoting the expression and mineralization of bone genes, can effectively solve the problems of poor infection control capability and insufficient bone integration capability of the existing skull repairing materials based on titanium mesh, polyether ether ester, polymethyl methacrylate and the like, and has unique advantages and good market application prospect when being used as a biological repairing material.
Description
Technical Field
The invention belongs to the field of skull biomaterials and tissue engineering, and particularly relates to a bioactive SBS material for skull repair and a preparation method and application thereof.
Background
Skull coloboma is easy to be caused by intracranial malignant tumor, traumatic brain injury, large-area cerebral infarction or intracranial excision caused by nervous diseases such as apoplexy and congenital skull deformity, and the brain tissue is not protected. And if the patient is not treated by cranioplasty after craniotomy, serious complications such as nerve dysfunction, brain deformation and fatal cerebrospinal fluid leakage are easy to occur, which seriously affects the life of the patient. Therefore, after intracranial resection, skull repair is usually required, and the reconstruction of skull defect is accompanied with the difficulty.
The autograft material mainly comprises autogenous skull and bone flap of the rest parts of human body, and is considered as the most suitable forming material for cranioplasty. However, when the bone flap is cut from the ilium, rib, sternum and other parts of the human body, the disease of the supply area can be caused, and the problem of secondary injury is limited. Moreover, since autologous bone is difficult to preserve and ethical after being taken out of the body, autologous bone flap is not preferable.
In contrast, xenosynthetic materials have received much attention as artificial craniums due to their advantages of short surgical time, no donor site morbidity, easy availability, good batch consistency, and no ethical issues. Currently, titanium mesh, Hydroxyapatite (HA), alumina ceramics, Polymethylmethacrylate (PMMA) and Polyetheretherketone (PEEK) are common synthetic skull repair materials. Although these materials have been successfully used in cranioplasty, there are still some problems in clinical practice. Poor infection control and inadequate osseointegration are major problems. Other disadvantages, such as high cost, high 3D printing temperature (for PEEK), thermal conductivity (for titanium mesh), brittleness (for HA and alumina ceramics), and exothermic combustion reaction (for PMMA) also hamper their development to some extent.
At the same time, it is noteworthy that there are 2 major complications after cranioplasty, one of which is surgical site infection, with an infection rate of up to 33%. In general, infection is highly correlated with the formation of biofilms. Biofilms are an aggregated group of bacteria adhering to the surface of wounds or implants and are 10 to 1000 times more resistant to antibiotics than planktonic bacteria, which is the key point for the difficulty in treating diseases associated with biofilm infections. Therefore, the inherent antimicrobial properties are critical for the implant to prevent infection and control early infection. Secondly, osteointegration is poor, with an increased risk of displacement closely related thereto. Thus, there is a need to develop a SBS skull forming material with bioactivity.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the bioactive SBS material for repairing the skull as well as the preparation method and the application thereof.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
a preparation method of a bioactive SBS material for skull restoration comprises the following steps: covalently connecting aldehyde-modified quaternary ammonium salt chitosan on a SBS (poly (styrene-butadiene-styrene)) substrate, and then fixing a polydopamine-modified zeolite imidazole ester framework-8 and polydopamine-modified hydroxyapatite on the surface of the chitosan.
Further, the preparation method of the quaternary ammonium salt chitosan comprises the following steps:
5g CS (chitosan) was dissolved in 180mL deionized water, followed by the dropwise addition of 900. mu.L of acetic acid solution (0.5% v/v) and stirring at 55 ℃ for 1 h; adding 5795 μ L of glycidol trimethyl ammonium chloride dropwise into the mixed solution, and reacting at 55 deg.C for 18 h; after the reaction is finished, centrifuging the solution at the speed of 4500r/min for 20min, then filtering the supernatant, and precipitating the filtrate in ethanol/acetone mixed solution (1:1, v/v); finally, the precipitate was dried in a vacuum oven at room temperature for 3d to obtain QCS (quaternary ammonium chitosan).
Further, the quaternary ammonium salt chitosan is modified by adopting a natural aldehyde compound.
Further, the natural aldehyde compound is protocatechuic aldehyde, perillaldehyde or vanillin and the like, and the modification process is as follows:
0.5g of QCS was weighed out and dissolved in 40mL of MES solution (pH 5.5) and stirred overnight; dissolving 1g protocatechualdehyde in 30mL of water/methanol mixed solution (2:1, v/v), stirring at room temperature for 3.5h, and then adding the solution into the QCS solution; 250mg of NaBH 4 Slowly adding into the mixed solution until no bubbles are generated, adjusting the pH value of the solution to 5.5, and stirring overnight; after the reaction is finished, the obtained product is put into a dialysis bag (MWCO 3500), and dialyzed for 2d in an acidic aqueous solution (pH 5.0, HCl), and finally, the QCSC (protocatechuic aldehyde modified quaternary ammonium salt chitosan) can be obtained by freeze drying.
Further, the process of covalently linking the modified quaternary ammonium salt chitosan on the SBS substrate is as follows:
(1) putting the SBS substrate into a silane coupling agent, performing plasma surface treatment and then cleaning to prepare the SBS-Si substrate;
(2) and (2) placing the SBS-Si substrate in the protocatechuic aldehyde modified quaternary ammonium salt chitosan solution, and soaking for 30-50 h at 50-70 ℃.
Further, the silane coupling agent is trimethoxysilane (GPTMS), KH560 or the like.
Further, the concentration of the protocatechuic aldehyde modified quaternary ammonium salt chitosan solution is 0.5-2 mg/mL.
Further, the SBS substrate was made from SBS filaments with a diameter of 1.75mm by 3D printing.
Further, the 3D printer used during 3D printing is a self-made device, the temperature of a spray head of the 3D printer is 210 ℃, and the temperature of a bed of the 3D printer is 110 ℃.
Further, the preparation method of the polydopamine modified zeolite imidazole ester framework-8 comprises the following steps:
adding 2-methylimidazole into a zinc acetate solution, adding dopamine hydrochloride, stirring for reacting for 4-6 hours, centrifuging at a speed of 10000-14000 r/min for 10-15 min, collecting and washing a solid-phase product, and drying at 60-80 ℃.
Further, the final concentration of the zinc acetate solution 2-methylimidazole is 1-2M; the final concentration of the dopamine hydrochloride after addition is 0.5-2 mg/mL.
Further, the preparation process of the polydopamine modified hydroxyapatite comprises the following steps:
(1) mixing Ca (NO) 3 ) 2 ·4H 2 O solution and (NH) 4 ) 2 HPO 4 Mixing the solutions, adjusting the pH value to 10, and stirring and reacting at 70-80 ℃ for 4-6 h; standing for 24 hours after the reaction is finished, collecting and washing a solid-phase product, filtering, freeze-drying for 20-30 hours, calcining for 2-5 hours at 300-400 ℃, and finally screening by using a 100-200-mesh sieve to obtain nano-hydroxyapatite;
(2) mixing the nano-hydroxyapatite solution with the dopamine hydrochloride solution, stirring for 4-6 h, centrifuging for 10-15 min at the speed of 10000-14000 r/min, collecting and washing a solid-phase product, and drying at 60-80 ℃.
Further, the volume ratio of the nano hydroxyapatite solution to the dopamine hydrochloride solution is 1: 1; wherein the content of the nano-hydroxyapatite in the nano-hydroxyapatite solution is 10-15 mg/mL, and the content of the dopamine hydrochloride in the dopamine hydrochloride solution is 10-15 mg/mL.
Further, Ca (NO) 3 ) 2 ·4H 2 O solution and (NH) 4 ) 2 HPO 4 The concentration of the solution was 0.5M, and the volume ratio of the two solutions was 1.67.
The bioactive SBS material for skull restoration prepared by the method.
The application of the bioactive SBS material in preparing cranial bone forming material.
The invention has the beneficial effects that:
1. the skull repairing material substrate is prepared from poly (styrene-butadiene-styrene), and the problems of high price, impact resistance, thermal impact of environment temperature on brain and the like of the skull repairing material based on titanium mesh, polyether ether ester and polymethyl methacrylate can be effectively solved.
2. According to the invention, protocatechuic aldehyde modified quaternary ammonium salt chitosan, a polydopamine modified zeolite imidazole ester framework and polydopamine modified hydroxyapatite are used as raw materials, wherein the protocatechuic aldehyde modified quaternary ammonium salt chitosan endows the material with good bactericidal performance; the polydopamine modified zeolite imidazole ester skeleton and the polydopamine modified hydroxyapatite are components for promoting the antibacterial property, the bone differentiation and the mineralization of the material.
Drawings
FIG. 1 is a photograph of SBS, SBS-QCSC, pHA/pZIF-8@ SBS-QCSC substrates;
FIG. 2 is an infrared spectrum of SBS, SBS-QCSC, pHA/pZIF-8@ SBS-QCSC substrates;
FIG. 3 is a SEM experimental result chart of the antibacterial activity of the bioactive SBS material;
FIG. 4 is a graph showing the results of experiments on the ability of the SBS material to promote bone differentiation.
Detailed Description
The following description of the embodiments of the present invention is provided to facilitate the understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and it will be apparent to those skilled in the art that various changes may be made without departing from the spirit and scope of the invention as defined and defined in the appended claims, and all matters produced by the invention using the inventive concept are protected.
Example 1
A preparation method of a bioactive SBS skull forming material comprises the following steps:
(1) preparation of protocatechuic aldehyde modified quaternary ammonium salt chitosan
S1, 5g of CS was dissolved in 180mL of deionized water, followed by dropwise addition of 900. mu.L of an acetic acid solution (0.5% v/v), and stirring was carried out at 55 ℃ for 1 h. Then, 5795. mu.L of glycidyltrimethylammonium chloride was added dropwise to the above mixture and reacted at 55 ℃ for 18 hours. Then the mixed solution is centrifuged at the speed of 4500r/min for 20min, the obtained supernatant is filtered, and the filtered solution is precipitated in ethanol/acetone (1:1, v/v) mixed solution. Finally, the obtained precipitate is dried in a vacuum drying oven at room temperature for 3d to obtain QCS.
S2, 500mg QCS was weighed out and dissolved in 40mL MES solution (pH 5.5) and stirred overnight. 1g protocatechualdehyde was dissolved in 30mL of a water/methanol (2:1, v/v) mixture and stirred at room temperature for 3.5h, then the solution was added to the above QCS solution. 250mg of NaBH 4 Slowly add to the mixture until no air bubbles are generated. The solution was adjusted to pH 5.5 and stirred overnight. After the reaction is finished, the obtained product is put into a dialysis bag (MWCO 3500) and is continuously dialyzed for 2d in an acidic aqueous solution (pH 5.0, HCl), and finally, the QCSC can be obtained by freeze drying.
(2) Preparation of Polydopamine modified Zeolite Imidazol ester framework-8
2-methylimidazole (2M) was added to a zinc acetate solution (0.08M) using Tris-buffer (10mM, pH 8.5) as a solvent, and then dopamine hydrochloride was immediately added to the mixture at a final concentration of 1mg/mL, and reacted for 4h with stirring. The resulting pZIF-8 suspension was centrifuged at 10000r/min for 10 min. And then washing the obtained precipitate for 3 times, and drying at 60 ℃ for 12h to obtain the pZIF-8 nano-particles.
(3) Preparation of Polydopamine modified hydroxyapatite
S1, Ca (NO) concentration of 0.5M 3 ) 2 ·4H 2 O solution and (NH) 4 ) 2 HPO 4 The solution was mixed at a volume ratio of 1.67 and NH was used 3 ·H 2 O the pH of the mixture was adjusted to 10, and the reaction was stirred at 70 ℃ for 4 hours. After the reaction was completed, the suspension was allowed to stand for 24 hours, and the obtained precipitate was washed 3 times, filtered, and freeze-dried for 24 hours. Subjecting the precipitate to a temperature of 300 deg.CCalcining for 2h, and screening by using a 200-mesh sieve to obtain the nano-hydroxyapatite.
S2, 10mL of Tris-buffer (10mM) containing nano-hydroxyapatite (100mg) prepared above and 10mL of Tris-buffer (10mM) containing dopamine hydrochloride (100mg) were mixed and stirred for 4h to obtain a pHA suspension. The suspension was centrifuged at 10000r/min for 10min, and the resulting precipitate was washed 3 times and then dried at 60 ℃ for 12h to obtain the pHA nanoparticles.
(4) Preparation of SBS substrates
SBS was prepared as a 1.75mm diameter filament and then processed by 3D printing. And washing the obtained printing product with ethanol for 3 times, and drying at 40 ℃ for 2 hours to obtain the 3D printed SBS substrate.
(5) Preparation of bioactive SBS material
S1, the SBS substrate (21 mm. times.21 mm. times.1 mm) was placed in 100. mu.L of GPTMS and treated with a plasma cleaner for 7 min. And washing the treated substrate with ethanol and water alternately for 3 times to obtain the SBS-Si substrate. QCSC was dissolved in MES solution at a final concentration of 1 mg/mL. Then soaking the SBS-Si substrate in the QCSC solution (pH 5.5) for 36h at 55 ℃ to obtain the SBS-QCSC substrate.
S2, the SBS-QCSC substrates were soaked in the pHA suspension for 40min, followed by soaking in the pZIF-8 suspension for 40min, during which the SBS-QCSC substrates were washed 3 times with Tris-buffer (10 mM). And repeating the steps for 3 times, and washing the substrate for the last time to obtain the bioactive SBS skull forming material (pHA/pZIF-8@ SBS-QCSC substrate).
Example 2
A preparation method of a bioactive SBS skull forming material comprises the following steps:
(1) preparation of protocatechuic aldehyde modified quaternary ammonium salt chitosan
S1, 5g of CS was dissolved in 180mL of deionized water, followed by dropwise addition of 900. mu.L of an acetic acid solution (0.5% v/v), and stirring was carried out at 55 ℃ for 1 h. Then, 5795. mu.L of glycidyltrimethylammonium chloride was added dropwise to the above mixture and reacted at 55 ℃ for 18 hours. Then the mixed solution is centrifuged at the speed of 4500r/min for 20min, the obtained supernatant is filtered, and the filtered solution is precipitated in ethanol/acetone (1:1, v/v) mixed solution. Finally, the obtained precipitate is dried in a vacuum drying oven at room temperature for 3d to obtain QCS.
S2, 500mg QCS was weighed out and dissolved in 40mL MES solution (pH 5.5) and stirred overnight. 1g protocatechualdehyde was dissolved in 30mL of a water/methanol (2:1, v/v) mixture and stirred at room temperature for 3.5h, then the solution was added to the above QCS solution. 250mg of NaBH 4 Slowly add to the mixture until no air bubbles are generated. The solution was adjusted to pH 5.5 and stirred overnight. After the reaction is finished, the obtained product is put into a dialysis bag (MWCO 3500) and is continuously dialyzed for 2d in an acidic aqueous solution (pH 5.0, HCl), and finally, the QCSC can be obtained by freeze drying.
(2) Preparation of Polydopamine modified Zeolite Imidazol ester framework-8
2-methylimidazole (2M) was added to a zinc acetate solution (0.08M) using Tris-buffer (10mM, pH 8.5) as a solvent, and then dopamine hydrochloride was immediately added to the mixture at a final concentration of 0.5mg/mL, and reacted for 4h with stirring. The resulting pZIF-8 suspension was centrifuged at 10000r/min for 10 min. And then washing the obtained precipitate for 3 times, and drying at 60 ℃ for 12h to obtain the pZIF-8 nano-particles.
(3) Preparation of Polydopamine modified hydroxyapatite
S1, Ca (NO) concentration of 0.5M 3 ) 2 ·4H 2 O solution and (NH) 4 ) 2 HPO 4 The solution was mixed at a volume ratio of 1.67 and NH was used 3 ·H 2 O the pH of the mixture was adjusted to 10, and the reaction was stirred at 70 ℃ for 4 hours. After the reaction was completed, the suspension was allowed to stand for 24 hours, and the obtained precipitate was washed 3 times, filtered, and freeze-dried for 24 hours. Calcining the precipitate at 300 ℃ for 2h, and screening by using a 200-mesh sieve to obtain the nano-hydroxyapatite.
S2, 10mL of Tris-buffer (10mM) containing nano-hydroxyapatite (100mg) prepared as above and 10mL of Tris-buffer (10mM) containing dopamine hydrochloride (150mg) were mixed and stirred for 4h to obtain a pHA suspension. The suspension was centrifuged at 10000r/min for 10min, and the resulting precipitate was washed 3 times and then dried at 60 ℃ for 12h to obtain the pHA nanoparticles.
(4) Preparation of SBS substrates
SBS was prepared as a 1.75mm diameter filament and then processed by 3D printing. And washing the obtained printing product with ethanol for 3 times, and drying at 40 ℃ for 2 hours to obtain the 3D printed SBS substrate.
(5) Preparation of bioactive SBS material
S1, the SBS substrate (21 mm. times.21 mm. times.1 mm) was placed in 100. mu.L of GPTMS and treated with a plasma cleaner for 7 min. And washing the treated substrate with ethanol and water alternately for 3 times to obtain the SBS-Si substrate. QCSC was dissolved in MES solution at a final concentration of 1.5 mg/mL. Then soaking the SBS-Si substrate in the QCSC solution (pH 5.5) for 36h at 55 ℃ to obtain the SBS-QCSC substrate.
S2, the SBS-QCSC substrates were soaked in the pHA suspension for 40min, followed by soaking in the pZIF-8 suspension for 40min, during which the SBS-QCSC substrates were washed 3 times with Tris-buffer (10 mM). And repeating the steps for 3 times, and washing the substrate for the last time to obtain the bioactive SBS skull forming material (pHA/pZIF-8@ SBS-QCSC substrate).
Example 3
A preparation method of a bioactive SBS skull forming material comprises the following steps:
(1) preparation of perillaldehyde modified quaternary ammonium salt chitosan
S1, 5g of CS was dissolved in 180mL of deionized water, followed by addition of 900. mu.L of acetic acid solution (0.5% v/v) dropwise and stirring at 55 ℃ for 1 h. Then, 5795. mu.L of glycidyltrimethylammonium chloride was added dropwise to the above mixture and reacted at 55 ℃ for 18 hours. Then the mixed solution is centrifuged at the speed of 4500r/min for 20min, the obtained supernatant is filtered, and the filtered solution is precipitated in ethanol/acetone (1:1, v/v) mixed solution. Finally, the obtained precipitate is dried in a vacuum drying oven at room temperature for 3d to obtain QCS.
S2, 500mg QCS was weighed out and dissolved in 40mL MES solution (pH 5.5) and stirred overnight. Perilla aldehyde (1 g) was dissolved in 30mL of a water/methanol (2:1, v/v) mixture and stirred at room temperature for 3.5h, and then the solution was added to the QCS solution.250mg of NaBH 4 Slowly add to the mixture until no air bubbles are generated. The solution was adjusted to pH 5.5 and stirred overnight. After the reaction is finished, the obtained product is put into a dialysis bag (MWCO 3500) and is continuously dialyzed for 2d in an acidic aqueous solution (pH 5.0, HCl), and finally, the QCSC can be obtained by freeze drying.
(2) Preparation of Polydopamine modified Zeolite Imidazol ester framework-8
2-methylimidazole (2M) was added to a zinc acetate solution (0.08M) using Tris-buffer (10mM, pH 8.5) as a solvent, and then dopamine hydrochloride was immediately added to the mixture at a final concentration of 1.5mg/mL and reacted for 4 hours with stirring. The resulting pZIF-8 suspension was centrifuged at 10000r/min for 10 min. And then washing the obtained precipitate for 3 times, and drying at 60 ℃ for 12h to obtain the pZIF-8 nano-particles.
(3) Preparation of Polydopamine modified hydroxyapatite
S1, mixing Ca (NO) at a concentration of 0.5M 3 ) 2 ·4H 2 O solution and (NH) 4 ) 2 HPO 4 The solution was mixed at a volume ratio of 1.67 and NH was used 3 ·H 2 O the pH of the mixture was adjusted to 10, and the reaction was stirred at 70 ℃ for 4 hours. After the reaction was completed, the suspension was allowed to stand for 24 hours, and the obtained precipitate was washed 3 times, filtered, and freeze-dried for 24 hours. Calcining the precipitate at 300 ℃ for 2h, and screening by using a 200-mesh sieve to obtain the nano-hydroxyapatite.
S2, 10mL of Tris-buffer (10mM) containing nano-hydroxyapatite (150mg) prepared as above and 10mL of Tris-buffer (10mM) containing dopamine hydrochloride (100mg) were mixed and stirred for 4h to obtain a pHA suspension. The suspension was centrifuged at 10000r/min for 10min, and the resulting precipitate was washed 3 times and then dried at 60 ℃ for 12h to obtain the pHA nanoparticles.
(4) Preparation of SBS substrates
SBS was prepared as a 1.75mm diameter filament and then processed by 3D printing. And washing the obtained printing product with ethanol for 3 times, and drying at 40 ℃ for 2 hours to obtain the 3D printed SBS substrate.
(5) Preparation of bioactive SBS material
S1, the SBS substrate (21 mm. times.21 mm. times.1 mm) was placed in 100. mu.L KH560 and treated with a plasma cleaner for 7 min. And washing the treated substrate with ethanol and water alternately for 3 times to obtain the SBS-Si substrate. QCSC was dissolved in MES solution at a final concentration of 1 mg/mL. Then soaking the SBS-Si substrate in the QCSC solution (pH 5.5) for 36h at 55 ℃ to obtain the SBS-QCSC substrate.
S2, the SBS-QCSC substrate was soaked in the pHA suspension for 40min, followed by soaking in the pZIF-8 suspension for 40min, during which the SBS-QCSC substrate was washed 3 times with Tris-buffer (10 mM). And repeating the steps for 3 times, and washing the substrate for the last time to obtain the bioactive SBS skull forming material (pHA/pZIF-8@ SBS-QCSC substrate).
Experimental example 1
The bioactive SBS material prepared in the embodiment 1 of the application is subjected to antibacterial detection, and the specific operation steps are as follows:
sterilizing SBS, SBS-QCSC, pHA/pZIF-8@ SBS-QCSC material (7mm × 7mm × 1mm) with 75% anhydrous ethanol, and further soaking in 200mL of bacterial suspension (2 × 10mm) 7 CFU/mL) and incubated at 37 ℃ for 48 h. The material was then removed and washed with PBS before bacterial fixation and SEM observation. The results of the experiment are shown in FIG. 3.
As can be seen from FIG. 3, both Pseudomonas aeruginosa and Staphylococcus aureus grew in clusters on SBS material and formed biofilms. Compared with SBS materials, the bacterial number of the surfaces of SBS-QCSC and pHA/pZIF-8@ SBS-QCSC materials is obviously reduced, which shows that the materials have good antibacterial property, and the antibacterial property of the pHA/pZIF-8@ SBS-QCSC materials is particularly obvious. Compared with the existing skull forming material, the prepared pHA/pZIF-8@ SBS-QCSC skull forming material has inherent antibacterial property.
Experimental example 2
The bioactive SBS material prepared in the embodiment 1 of the application is subjected to osteogenic differentiation capacity detection, and the specific operation steps are as follows:
sterilizing SBS, SBS-QCSC, pHA/pZIF-8@ SBS-QCSC material (7mm × 7mm × 1mm) with 75% anhydrous ethanol, placing in 48-well plate, and further sterilizingMC3T3-E1 cells at 5X 10 4 The density of individual cells/well is seeded onto the material and cultured. The medium was complete medium containing 50. mu.g/mL L-ascorbic acid and 10mM disodium beta-glycerophosphate hydrate, exchanged every 2 d. Used after culturing for 7, 14 and 21d
(Thermo Fisher Scientific, USA) Total RNA was extracted from MC3T3-E1 cells on each substrate. The extracted total RNA was then reverse transcribed using the reverse transcription System kit (Thermo Fisher Scientific, USA), and finally RT-qPCR sequenced using 2 -ΔΔCt The method processes the data to analyze the mRNA expression level. The results of the experiment are shown in FIG. 4.
As can be seen from fig. 4, MC3T3-E1 cells cultured with the pHA/pZIF-8@ SBS-QCSC material exhibited significantly increased levels of expression of alkaline phosphatase (ALP), Runt-associated transcription factor 2(Runx2), and collagen type I (Col I) compared to MC3T3-E1 cells cultured with SBS and SBS-QCSC materials. ALP, Runx2 and Col I are osteogenic differentiation related genes, the higher the expressed mRNA level is, the stronger the osteogenic differentiation capacity of cells is, and the further indication that the prepared pHA/pZIF-8@ SBS-QCSC skull forming material has obvious advantages in promoting the osteogenic gene expression.
Claims (10)
1. A preparation method of a bioactive SBS material for skull restoration is characterized by comprising the following steps: covalently connecting the modified quaternary ammonium salt chitosan on an SBS substrate, and then fixing a polydopamine modified zeolite imidazole ester skeleton-8 and polydopamine modified hydroxyapatite on the surface of the modified quaternary ammonium salt chitosan.
2. The preparation method according to claim 1, wherein the quaternary ammonium salt chitosan is modified by natural aldehyde compounds, and the modification process comprises the following steps:
dissolving quaternary ammonium salt chitosan, adding natural aldehyde compound solution, and then adding NaBH 4 And adjusting the pH value of the solution to 5.5, stirring overnight, and dialyzing in an acid environment for 2-3 days.
3. The method according to claim 2, wherein the natural aldehyde compound is protocatechuic aldehyde, vanillin, or perillaldehyde.
4. The method according to claim 3, wherein the covalent bonding of the modified quaternary ammonium salt chitosan on the SBS substrate is performed by:
(1) putting the SBS substrate into a silane coupling agent, performing plasma surface treatment and then cleaning to prepare the SBS-Si substrate;
(2) and (2) placing the SBS-Si substrate in a natural aldehyde modified quaternary ammonium salt chitosan solution, and soaking for 30-50 h at 50-70 ℃.
5. The method according to claim 1, wherein the polydopamine-modified zeolitic imidazolate framework-8 is prepared by:
adding 2-methylimidazole into a zinc acetate solution, adding dopamine hydrochloride, stirring for reacting for 4-6 hours, centrifuging for 10-15 minutes at a speed of 10000-14000 r/min, collecting and washing a solid-phase product, and drying at 60-80 ℃.
6. The preparation method according to claim 5, wherein the final concentration of 2-methylimidazole in the zinc acetate solution is 1-2M; the final concentration of the dopamine hydrochloride after addition is 0.5-2 mg/mL.
7. The preparation method according to claim 1, wherein the polydopamine-modified hydroxyapatite is prepared by the following steps:
(1) mixing Ca (NO) 3 ) 2 ·4H 2 O solution and (NH) 4 ) 2 HPO 4 Mixing the solutions, adjusting the pH value to 10, and stirring and reacting at 70-80 ℃ for 4-6 h; standing for 24 hours after the reaction is finished, collecting and washing a solid-phase product, filtering, freeze-drying for 20-30 hours, calcining for 2-5 hours at 300-400 ℃, and finally screening by using a 100-200-mesh sieve to obtain the productObtaining nano hydroxyapatite;
(2) mixing the nano-hydroxyapatite solution with the dopamine hydrochloride solution, stirring for 4-6 h, centrifuging for 10-15 min at the speed of 10000-14000 r/min, collecting and washing a solid-phase product, and drying at 60-80 ℃.
8. The preparation method according to claim 7, wherein the volume ratio of the nano-hydroxyapatite solution to the dopamine hydrochloride solution is 1: 1; wherein the content of the nano-hydroxyapatite in the nano-hydroxyapatite solution is 10-15 mg/mL, and the content of the dopamine hydrochloride in the dopamine hydrochloride solution is 10-15 mg/mL.
9. The bioactive SBS material prepared by the method of any one of claims 1-8 and used for skull restoration.
10. Use of the bioactive SBS material for skull repair according to claim 9 in the preparation of bone biomaterials, bone tissue engineering materials or skull forming materials.
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| CN115721776A (en) * | 2022-11-10 | 2023-03-03 | 奥精医疗科技股份有限公司 | Preparation method of children skull repairing material |
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| EP2338531A1 (en) * | 2009-12-28 | 2011-06-29 | AKADEMIA GORNICZO-HUTNICZA im. Stanislawa Staszica | Method for fabrication of synthetic bioceramic implant material based on carbonate hydroxyapatites |
| CN114504677A (en) * | 2022-01-11 | 2022-05-17 | 武汉亚洲生物材料有限公司 | 3D printing skull repairing titanium mesh and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP2338531A1 (en) * | 2009-12-28 | 2011-06-29 | AKADEMIA GORNICZO-HUTNICZA im. Stanislawa Staszica | Method for fabrication of synthetic bioceramic implant material based on carbonate hydroxyapatites |
| CN114504677A (en) * | 2022-01-11 | 2022-05-17 | 武汉亚洲生物材料有限公司 | 3D printing skull repairing titanium mesh and preparation method thereof |
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| CN115721776A (en) * | 2022-11-10 | 2023-03-03 | 奥精医疗科技股份有限公司 | Preparation method of children skull repairing material |
| CN115721776B (en) * | 2022-11-10 | 2024-02-20 | 奥精医疗科技股份有限公司 | Preparation method of skull repairing material for children |
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