摘要
目的探讨硫化铜(CuS)/氧化石墨烯(GO)/壳聚糖(CS)/纳米羟基磷灰石(nHA)复合材料(CGCHs)的抗菌和促成骨作用及其作用机制。方法采用水热法合成CuS/GO纳米颗粒,通过原位沉淀法合成CS/nHA支架和CGCHs支架,检测材料表征、光热转换性能和生物安全性,评估CGCHs组和近红外光(NIR)照射下CGCHs(CGCHs+NIR)组的细菌抑制效果及其对细菌生物膜相关基因表达的影响,观察CGCHs和CS/nHA不同材料组的促成骨分化和成骨、破骨相关基因表达。结果CGCHs是具有高度孔隙率的三维支架,在CuS/GO浓度为200μg/mL时CGCHs同时兼具良好的红外升温效果和生物安全性。琼脂糖平板涂菌和细菌死活染色结果均表明CGCHs+NIR组抗菌性能最佳,生物膜相关基因qPCR检测证实其具有抑制细菌生物膜相关基因表达的作用。茜素红染色结果表明CGCHs具有良好的体外促成骨性能,体外共培养3、7、14、21和28 d qPCR结果表明CGCHs对成骨早期和晚期相关基因表达均具有促进作用。与破骨细胞共培养结果可观察到CGCHs具有抑制破骨细胞形成的作用,细胞凋亡检测结果进一步验证这一结论,破骨分化相关基因qPCR检测结果表明,CGCHs主要通过抑制抗酒石酸酸性磷酸酶、组织蛋白酶K、CTR、P65和P38在共培养7、14 d的表达来抑制破骨细胞的分化。结论作为纳米复合材料,CGCHs生物安全性好,具有良好的红外光热协同抗菌作用,在促成骨分化的同时抑制破骨细胞分化,有望为感染性骨缺损治疗提供新的思路。
Objective To investigate the antibacterial and bone-promoting effects of copper sulfide(CuS)/graphene oxide(GO)/chitosan(CS)/nano-hydroxyapatite(nHA)composite nanomaterials(CGCHs),and further explore the mechanisms of these two effects.Methods CuS/GO nanoparticles were synthesized by hydrothermal method,then CS/nHA scaffolds and CGCHs scaffolds were synthesized by in-situ precipitation method,the characterization,photothermal conversion performance and biosafety of the synthesized materials were tested,the antibacterial performance as well as the effects on the expression of bacterial biofilm-related genes were evaluated and compared between CGCHs group and CGCHs+NIR(CGCHs under near-infrared irradiation)group,and the features of promoting bone differentiation and inhibiting osteoclastic differentiation as well as the expressions of osteogenesis and osteoclast related genes were observed in CGCHs group and CS/nHA group.Results The material characterization test showed that CGCHs was a three-dimensional scaffold with high porosity,and had both good infrared heating effect and biosafety when CuS/GO concentration was 200μg/mL.The results of agarose plate coating and bacteria dead staining showed that CGCHs+NIR group had the best antibacterial performance,and qPCR detection of biofilm-related genes showed that CGCHs+NIR group also inhibited the expression of biofilm-related genes.The results of alizarin red staining revealed that CGCHs had good osteogenic activity in vitro,and qPCR results after 3,7,14,21,28 d co-culture in vitro showed that CGCHs could promote the expression of related genes in the early and late stages of osteogenesis.It was observed that CGCHs inhibited osteoclast formation in co-culture with osteoclasts,and the results of apoptosis further verified this conclusion.The dectection of qPCR of osteoclast differentiation-related genes proved that CGCHs inhibited osteoclast differentiation mainly by inhibiting the expression of tartrate resistant acid phosphatase,cathepsin K,CTR,P65 and P38 at 7 d and 14 d after co-culture.Conclusions CGCHs has good biosafety,and good infrared photothermal synergistic antibacterial effect,which can promote bone differentiation and inhibit osteoclastic differentiation at the same time.Therefore,CGCHs composite nanomaterials are expected to provide new ideas for the treatment of infectious bone defects.
作者
冯治滨
王万舜
陈玲玲
孔志远
张涛
马向阳
夏虹
FENG Zhibin;WANG Wanshun;CHEN Lingling;KONG Zhiyuan;ZHANG Tao;MA Xiangyang;XIA Hong(Department of Orthopedics,Suining Central Hospital,Suining,Sichuan 629000,China;不详)
出处
《中国骨科临床与基础研究杂志》
2024年第1期32-46,共15页
Chinese Orthopaedic Journal of Clinical and Basic Research
关键词
骨代用品
硫化铜
氧化石墨烯
壳聚糖
羟基磷灰石类
纳米复合物
谱学
近红外线
细胞毒性
消炎药
微生物敏感性试验
基因表达
骨生成
破骨细胞
Bone substitute
Copper sulfide
Graphene oxide
Chitosan
Hydroxyapatites
Nanocomposites
Spectroscopy,near-infrared
Cytotoxicity
Anti-inflammatory agents
Microbial sensitivity tests
Genetic expression
Osteogenesis
Osteoclasts
