硫化铜/氧化石墨烯/壳聚糖/纳米羟基磷灰石复合材料的抗菌及促成骨作用  

Antimicrobial and bone-enhancing effects of copper sulfide/graphene oxide/chitosan/nano-hydroxya patite composite nanomaterials

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)

机构地区：[1]遂宁市中心医院骨科,四川629000 [2]广州中医药大学第二临床医学院,510405 [3]中国人民解放军南部战区总医院骨科,广州510010

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

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

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 

分 类 号：R318.08[医药卫生—生物医学工程] R336[医药卫生—基础医学]