Table 2.
Silencing experiments of different siRNA nanosystems in vitro and vivo
| Delivery system | In vitro cell model | In vitro gene silencing result | Animal model | In vivo dose and administration interval | In vivo gene silencing result | Ref |
|---|---|---|---|---|---|---|
| Lipid-based nanoparticles | ||||||
| Neutral liposome–hyaluronan–integrin mAb | TK-1 cells | Suppression of CyD1, proinflammatory TH1 cytokine of IFN-γ, IL-2, IL-12, TNF-α. | C57BL/6 mice with DSS-induced | CyD1-siRNA 2.5 mg/kg entrapped in β7 I-tsNPs at days 0, 2, 4, and 6 | Reduction in mRNA expression of CyD1 and IL-12, TNF-α. Drastic suppression of intestinal damage, leukocyte infiltration, reversal body weight loss. | 30 |
| Polysaccharide-based nanoparticles | ||||||
| Modified chitosan–UAC–PEG–scCD98 | RAW 264.7 cells and colon-26 cells with LPS-induced | CD98 expression decreases to 27% of the control level. | Chronic colitis: transfer wild-type CD4+CD45RBhigh T-cells into RAG1−/− mice. Acute colitis: male C57BL/6 mice with DSS-induced |
Chronic colitis: 1 mg/kg of CD98 siRNA-loaded NPs double gavage twice a week for 5 consecutive weeks. Acute colitis: 2 mg/kg of CD98 siRNA-loaded NPs double gavage twice daily for 4 consecutive days | Chronic colitis: mRNA expression of CD98, TNF-α, IL-6, and IL-12 was decreased to 65.0%, 59.9%, 80.4%, and 31.8%, respectively, compared with control. Significant reduction of weight loss and MPO (~65.7%). Acute colitis: mRNA expression of CD98, TNF-α, IL-6, and IL-12 was decreased to 47.7%, 26.0%, 81.2%, and 71.2%, respectively. Body weight and MPO activity were correspondingly decreased. | 41 |
| Galactosylated trimethyl chitosan–cysteine | RAW 264.7 cells with LPS-induced | Expressions of Map4k4 and TNF-α mRNA were sharply decreased by 79.9% and 78.9%. | Male C57BL/6 mice with DSS-induced | SiRNA dose of 250 mg/kg per day for 6 consecutive days | Expressions of Map4k4 and TNF-α mRNA were sharply decreased by 92.1% and 69.0%. Inhibition of MPO activity, body weight loss, and colon shortening. | 46 |
| SC12-cyclodextrin-click-propylamine | RAW 264.7 cells with LPS-induced | TNF-α and IL-6 mRNA drop ~21- and sevenfold compared to control. | Female C57BL/6JOlaHsD mice | The volume of 100 µL solution administered to mice on day 2 and day 4 post-DSS treatment | Proximal colon tissue shows more significant TNF-α and IL-6 silencing than distal colon. Drastic reduction of TNF-α and IL-6 mRNA expression by 73%±13% and 58%±19%. | 57 |
| β-1,3-d-glucan | Peritoneal exudate cells with LPS-induced isolated from C57BL6/J male mice | TNF mRNA and TNF-α protein were inhibited by 50% and 30%. | C57BL6/J mice | 20 mg siRNA/kg | NPs increase survival rates of model mice. Reduction of TNF-α and IL-1β accompanied with TNF-α silencing. | 59 |
| PLA-based nanoparticles | ||||||
| PLA | RAW 264.7 cells with LPS-induced | Significant reduction of TNF-α (175.9 vs 559.8 pg/mL). | C57BL/6 mice | The dose of 5 mg NPs/mL hydrogel pre-treated for 4 days before LPS treatment | Significantly TNF-α reduction in colonic tissue and blood. (Colon tissue 7.5 vs 136.2 pg/mL blood: 1,751.5 vs 2,084.5 pg/mL) | 63 |
| PLA | Embryonic fibroblasts in mice | Efficient suppression of Klf4 protein. | C57BL/6 mice | The dose of 0.5 ng siRNA in 1 mg NPs and 100 mL hydrogel per mice daily for 7 consecutive days | Reduction of clinical score and MPO activity in mice with colitis. | 65 |
| PLA–PEG–maleimide–Ab | RAW 264.7 cells with LPS-induced | Significant reduction of TNF-α and enhanced kinetics of uptake. | C57BL/6 mice | Daily gavages of hydrogel-encapsulated NPs (10 mg/mL) during the 7/8 days with DSS treatment | 60 µg/kg TNF-α siRNA silence, 60% TNF-α expression, and 30% NPs were uptaken in intestinal macrophages in vivo. Higher IKβα accumulation in the treatment group than control. | 66 |
| CaP/PLGA-based nanoparticles | ||||||
| CaP/PLGA/PEI | MODE-K | The expression of IP-10 gene was reduced 30% as well as the expression of TNF-α or KC gene was reduced 50%. | BALB/c mice | CaP/PLGA nanoparticles (12 µg) were administered intrarectally from days 2 to 5 after DSS treatment | The expression of TNF-α mRNA decreased 40% and the expression of KC and IP-10 decreased 50% in the colon tissue. | 72 |
| NiMOS-based microspheres | ||||||
| NiMOS | – | – | Female Balb/c mice | The dose of 1.2 mg/kg body weight in 200 µL final volume | Reduction of TNF-α, IL-1β, IFN-γ, and MCP-1. Increasing body weight and falling levels of MPO. | 80 |
| NiMOS | – | – | Female Balb/c mice | The dose of 1.2 mg/kg body weight | Reduction of TNF-α, IL-1, IFN-γ. Increasing body weight and falling levels of MPO. | 77 |
| Thioketal-based nanoparticles | ||||||
| PPADT | RAW 264.7 cells with LPS-induced | Significant reduction of TNF-α production. | C57BL/6 mice | The dose of 2.3 or 0.23 mg siRNA/kg per day for 6 consecutive days | Reduction of mRNA expression of TNF-α, IL-6, IL-1, and IFN-γ. | 83 |
| PEI-based nanoparticles | ||||||
| p(CBA–B-PEI)–PEG–Man | RAW 264.7 cells with LPS-induced | Reduction of TNF-α production and the effects depend on the weight ratio of TPP-PPM/siRNA | Colitis tissues from FVB male mice with DSS induced | NPs added to the wells contain culture medium and colonic tissues, final TNF-α siRNA concentration in the medium is set as 100, 200, and 300 nM | TNF-α expression was decreased 61.0%. NPs were taken up by 29.5% colon macrophages. | 90 |
Note: “–” Indicates data not available.
Abbreviations: Ab, antibody; B-PEI, branched-polyethylenimine; CaP, calcium phosphate; CyD1, cyclin D1; DSS, dextran sulfate sodium; IFN, interferon; IL, interleukin; Klf4, Krüppel-like factor 4; LPS, lipopolysaccharides; mAb, monoclonal antibody; Map4k4, mitogen-activated protein kinase kinase kinase kinase 4; Man, mannose; MCP, monocyte chemotactic protein; MPO, myeloperoxidase; NiMOS, nanoparticles-in-microsphere oral system; NPs, nanoparticles; PEG, polyethylene glycol; PLA, polylactide; PLGA, poly(d,l-lactide-co-glycolide acid); ScCD98, single-chain CD98; siRNA, short interfering RNA; TH, T-helper cell; UAC, urocanic acid.