The research on immune therapeutic strategy of dual-function cationic nano delivery system for adjunctive treatment of periodontitis
10.19405/j.cnki.issn1000–1492.2026.09.006
- VernacularTitle:双效阳离子纳米递送体系的免疫治疗策略用于牙周炎辅助治疗的研究
- Author:
Yumeng ZHU
1
;
An LIU
1
Author Information
1. College & Hospital of Stomatology， Anhui Medical University， Anhui Provincial Key Laboratory of Oral Diseases Research， Hefei 230032
- Publication Type:Journal Article
- Keywords:
periodontitis;
nanocarrier;
Bruton’s tyrosine kinase;
siRNA;
antibacterial and anti-inflammatory
- From:
Acta Universitatis Medicinalis Anhui
2026;61(9):1548-1557
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effects of a dual-functional cationic nucleic acid nano delivery system （DCNA） loaded with small interfering RNA targeting Bruton’s tyrosine kinase （siBTK） on the immune-inflammatory microenvironment of periodontitis by regulating BTK expression. MethodsDCNAsiBTK was prepared using a double-emulsion solvent evaporation method， and its physicochemical properties and sustained-release performance were characterized. Cytotoxicity was evaluated using the cell counting kit-8 （CCK-8） assay. Antibacterial activity against periodontal pathogens was assessed by colony-forming assays. BTK mRNA and protein expression levels were determined by reverse transcription quantitative real-time polymerase chain reaction （RT-qPCR） and Western blot， respectively. Enzyme-linked immunosorbent assay （ELISA） was performed to quantify the levels of inflammatory cytokines， including IL-1β， IL-6， and TNF-α. A murine periodontitis model was established and treated with DCNAsiBTK. Therapeutic efficacy and biosafety were evaluated using quantitative polymerase chain reaction （qPCR）， micro-computed tomography （micro-CT）， hematoxylin-eosin （HE） staining， and immunohistochemistry （IHC）. ResultsStable DCNAsiBTK nanoparticles with an average particle size of （168.8±2.2） nm and a zeta potential of （17.9±0.8） mV were successfully prepared. Transmission electron microscopy （TEM） revealed uniformly spherical morphology with favorable sustained-release properties. CCK-8 assays demonstrated no significant cytotoxicity of DCNAsiBTK within the concentration range of 0-100 nmol/L. In vitro studies showed that DCNAsiBTK effectively inhibited the growth of Porphyromonas gingivalis （Pg）， Aggregatibacter actinomycetemcomitans （Aa）， and Fusobacterium nucleatum （Fn）， indicating broad-spectrum antibacterial activity. RT-qPCR and Western blot analyses demonstrated that DCNAsiBTK downregulated BTK mRNA and protein expression in a dose-dependent manner. ELISA results showed that DCNAsiBTK significantly reduced the secretion of IL-1β， IL-6， and TNF-α， thereby attenuating inflammatory responses. In vivo experiments further demonstrated that DCNAsiBTK suppressed anaerobic bacterial proliferation within periodontal pockets， alleviated alveolar bone resorption and periodontal inflammation， and caused no obvious pathological damage in major organs， including heart， liver， spleen， lungs， and kidneys. ConclusionDCNAsiBTK efficiently encapsulated and released siBTK， and exhibited dual activities including antibacterial and anti-inflammatory. By suppressing BTK expression， DCNAsiBTK effectively attenuated the progression of periodontitis， providing a promising novel strategy for adjunctive periodontal therapy.
