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

FENG Zhibin; WANG Wanshun; CHEN Lingling; KONG Zhiyuan; ZHANG Tao; MA Xiangyang; XIA Hong

doi:10.3969/j.issn.1674-666X.2024.01.006

Abstract：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 200 μg/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.