| Clinical treatment of craniofacial bone defects,which are often caused by tumor,trauma and infection,is still a huge challenge.The development of bone tissue engineering brings hope for the repair and treatment of bone defects.Due to its similar composition to normal bone,good bone conductivity and biodegradability,β-tricalcium phosphate(β-TCP)has been widely used in orthopedic reconstructive surgery,and has become one of the basic materials commonly used in 3D printing,but its osteogenic induction ability is limited.Although traditional growth factors have good osteogenic induction performance,they may have some side effects,such as ectopic osteogenesis.Therefore,it has become a hot topic of medical research to find drugs with good osteogenic induction and few side effects.naringin(NG)is the main active monomer component of the traditional Chinese medicine Rhizoma Drynariae,belonging to dihydroflavonoids,mainly exists in citrus species,has anti-inflammatory,anti-tumor and prevention of osteoporosis and other effects.Previous studies have shown that naringin can induce bone mesenchymal stem cells(BMSCs)to differentiate into osteoblasts through the Wnt/β-catenin signaling pathway.Therefore,in this study,a β-TCP scaffold was prepared by 3D printing technology.Naringin was loaded to the β-TCP scaffold assisted by polydopamine(PDA)coating.A series of physical characterization,drug release,and cytocompatibility tests were conducted to explore its potential to become a bone tissue engineering scaffold.1.β-TCP scaffold(group T)was prepared by 3D printing technology.After PDA coating(group D),the scaffolds were immersed in naringin ethanol solution with concentrations of 0.1 mg/ml,1 mg/ml and 5 mg/ml,respectively.Three NG/PDA/β-TCP composite stents containing naringin in different concentrations were obtained(group L,group M and group H,respectively,from low to high concentration).Scanning electron microscopy,energy spectrum analyzer and X-ray diffraction analyzer were used to detect the microstructure,element distribution and phase of the scaffold.Then the pore size,porosity,mechanical properties,hydrophilicity and drug release were measured.The results showed that the five groups of scaffolds had interconnected macroporous and microporous structures with uniform pore size.The apertures of T,D,L,M and H groups were(432.14±11.27)μm,(431.92±10.22)μm,(430.36±10.17)μm,(432.16±10.01)μm,(431.98±10.14)μm,respectively,with no significant difference(p >0.05).The porosity was(50.17±0.95)%,(50.47±0.12)%,(49.94±0.72)%,(50.15±0.91)%,(50.09±0.28)%,with no significant difference(p > 0.05).The compression moduli were(25.5±5.3),(25.2±2.19),(25.6±3.48),(25.6±4.14),(25.3±6.19)MPa,respectively,with no significant difference(p > 0.05);All the five groups of scaffolds had good hydrophilicity.The four groups of scaffolds coated by PDA all contain N element,and the substrate phase does not change,the surface of the scaffolds is rougher than that of the T group,and a large number of fine particle structure can be seen.Naringin in the three groups was released quickly in the first 3 days,and then slowly.At 30 days,the total release of naringin in group L,Group M and group H was 0.217 mg,0.251 mg and 0.410 mg,respectively.2.The effects of the scaffold on the proliferation,toxicity,adhesion and ALP activity of MC3T3-E1 cells were detected by CCK8 assay,cell survival and death staining assay,cell adhesion assay,and alkaline phosphatase(ALP)staining.The results showed that there was no cytotoxicity in all groups,among which group H had the strongest ability to promote cell proliferation(p < 0.001),the best cell adhesion performance and the strongest ALP activity.This series of experiments confirmed that the 3D-printed NG/PDA/β-TCP composite scaffold loaded with high concentration(5mg/ml)naringin has good biocompatibility,and can promote the proliferation,adhesion and ALP activity of MC3T3-E1 cells,which is expected to be used in bone tissue engineering. |
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