Wu Shizhou, Liu Yunjie, Zhang Hui, Lei Lei
Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, China.
State Key Laboratory of Oral Diseases, Department of Preventive Dentistry, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
J Dent Sci. 2020 Mar;15(1):65-74. doi: 10.1016/j.jds.2019.09.006. Epub 2019 Oct 23.
BACKGROUND/PURPOSE: () is considered a predominant pathogen for persistent periapical infections. Antisense (AS) RNA was reported to inhibit the biofilm formation and sensitized to calcium hydroxide medication. The aims of this study were to investigate whether the graphene oxide (GO) nanosheets could be used to enhance antibacterial activity of AS RNA for in periapical periodontitis.
We developed a graphene-based plasmid transformation system by loading antisense plasmid with GO-polyethylenimine (PEI) complexes (GO-PEI-AS). The particle size distributions and zeta-potential of the GO-PEI-AS were evaluated. Then, AS plasmids were labeled with gene encoding enhanced green fluorescent protein (ASR-eGFP). The transformation efficiencies and the bacterial viability of were evaluated by confocal laser scanning microscopy. Quantitative real-time PCR assays were used to investigate the expressions of virulent genes after transformed by GO-PEI-AS. Also, the antibacterial properties of the GO-PEI-AS were validated in the rat periapical periodontitis model.
We showed that GO-PEI could efficiently deliver the AS plasmid into cell. GO-PEI-AS significantly reduced virulent-associated gene expressions. Furthermore, GO-PEI-AS suppressed biofilm aggregation and improved bactericidal effects using infected canal models . In four-weeks periapical infective rat models, the GO-PEI-AS strains remarkably reduced the periapical lesion size.
Transformation efficiency and antibacterial prosperity of AS can be marked improved by GO-PEI based delivery system for infections.
背景/目的：（某菌）被认为是持续性根尖周感染的主要病原菌。据报道，反义（AS）RNA可抑制生物膜形成，并使（该菌）对氢氧化钙药物敏感。本研究的目的是探讨氧化石墨烯（GO）纳米片是否可用于增强AS RNA对根尖周炎中（该菌）的抗菌活性。
我们通过将反义（某菌）质粒与GO-聚乙烯亚胺（PEI）复合物（GO-PEI-AS）加载，开发了一种基于石墨烯的质粒转化系统。评估了GO-PEI-AS的粒径分布和zeta电位。然后，用编码增强型绿色荧光蛋白的基因标记AS质粒（ASR-eGFP）。通过共聚焦激光扫描显微镜评估（该菌）的转化效率和细菌活力。采用定量实时PCR检测法研究GO-PEI-AS转化后（该菌）毒力基因的表达。此外，在大鼠根尖周炎模型中验证了GO-PEI-AS的抗菌性能。
我们发现GO-PEI可以有效地将AS质粒导入（该菌）细胞。GO-PEI-AS显著降低了毒力相关基因的表达。此外，GO-PEI-AS使用感染根管模型抑制了生物膜聚集并提高了杀菌效果。在四周的根尖周感染大鼠模型中，GO-PEI-AS菌株显著减小了根尖周病变的大小。
基于GO-PEI的递送系统可显著提高AS对（该菌）感染的转化效率和抗菌效果。
