Ghasemi Maryam, Bakhshi Bita, Khashei Reza, Soudi Sara
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Nanobiomedicine (Rij). 2022 Jun 3;9:18495435221088374. doi: 10.1177/18495435221088374. eCollection 2022 Jan-Dec.
The aim of this study was to evaluate the cytotoxicity and immune-stimulatory effect of Mesoporous silica nanoparticle (MSN) Nano-adjuvant on pro-inflammatory cytokines and pattern recognition receptors (PRR) genes expression in Caco-2/PBMC co-culture model. MSNs were synthesized and characterized by scanning electron microscope (SEM), Brunauer Emmett Teller (BET) and Barrett Joyner Halenda (BJH) techniques. The BET specific surface area of MSNs was around 947 m/g and the total pore volume and average pore diameter were 1.5 cm/g and 8.01 nm, respectively. At the concentration of 10 µg/mL, MSN showed a low and time-dependent cytotoxicity on Caco-2 cells, while no cytotoxic effect was observed for 0.1 and 1 µg/mL concentrations after 24, 48 and 72 h. The expression of pro-inflammatory cytokines genes (IL-1, IL-8 and TNF-α) in co-cultures treated with different concentrations of MSN showed a dose-dependent significant increase up to 17.44, 2.722 and 4.34 folds, respectively, while the expression augmentation of IL-1 gene was significantly higher than the others. This indicates slight stimulation of intestinal inflammation. Different concentrations of MSN significantly increased TLR4 and NOD2 expression to 4.14 and 2.14 folds, respectively. NOD1 was not affected significantly. It can be concluded that MSN might increase protective immune responses against antigens as a vaccine adjuvant candidate. It seems that stimulation of TNF-α, IL-1, and IL-8 expression in enterocytes probably transpires through the agonistic activity of MSN for TLRs including TLR4, while NOD2-associated signaling pathways are also involved. This study provides an overall picture of MSN as a novel and potent oral adjuvant for mucosal immunity.
本研究的目的是评估介孔二氧化硅纳米颗粒（MSN）纳米佐剂对Caco-2/PBMC共培养模型中促炎细胞因子和模式识别受体（PRR）基因表达的细胞毒性和免疫刺激作用。通过扫描电子显微镜（SEM）、布鲁诺尔-埃米特-泰勒（BET）和巴雷特-乔伊纳-哈伦达（BJH）技术合成并表征了MSN。MSN的BET比表面积约为947 m/g，总孔体积和平均孔径分别为1.5 cm/g和8.01 nm。在10 µg/mL浓度下，MSN对Caco-2细胞表现出低且随时间变化的细胞毒性，而在24、48和72小时后，0.1和1 µg/mL浓度未观察到细胞毒性作用。用不同浓度MSN处理的共培养物中促炎细胞因子基因（IL-1、IL-8和TNF-α）的表达分别呈剂量依赖性显著增加，最高可达17.44、2.722和4.34倍，而IL-1基因的表达增强明显高于其他基因。这表明肠道炎症受到轻微刺激。不同浓度的MSN分别将TLR4和NOD2的表达显著提高到4.14和2.14倍。NOD1未受到显著影响。可以得出结论，MSN作为疫苗佐剂候选物可能会增加针对抗原的保护性免疫反应。似乎MSN对包括TLR4在内的TLR的激动活性可能导致肠细胞中TNF-α、IL-1和IL-8表达的刺激，同时NOD2相关信号通路也参与其中。本研究提供了MSN作为新型强效口服黏膜免疫佐剂的全貌。
