The effect of the shape of mesoporous silica nanoparticles on cellular uptake and cell function

The Shape Effect of Mesoporous Silica Nanoparticles on Biodistribution, Clearance, and Biocompatibility in Vivo

2012.04.28

孙亚娟

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研究背景

研究方法

结论

研究结果

目录

Tianjin University 2012-04-28

研究背景

Nanoenabled drug delivery systems are gaining application in the pharmaceutical industry, since nanoparticle-based drugs may have improved solubility and altered pharmacokinetics and biodistributions compared to small molecule drugs.

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The design of smart functional nanosystems for intracellular imaging and therapeutic applications requires a thorough understanding of the mechanisms by which nanoparticles (NPs) enter cells.

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研究方法

Fig 1. Controlled fabrication and characterization of MSN-FITC. (A) Illustration of the fabrication of MSN-FITC by a specifically designed two-step preparation. (B) TEM images of different shaped MSN-FITC. The shapes of particles were controlled by CTAB concentration. (C) Relationship of CTAB concentration and aspect ratios (ARs). (D) Relationship of aqueous ammonia concentration and diameters.

Fig 2. Characterization of NSRFITC and NLR-FITC. (A) TEM image of NSRFITC. (B) TEM image showing the mesostructure of NSR-FITC. (C) TEM image of NLR-FITC. (D) TEM image showing the mesostructure of NLR-FITC. (E) Absorption and emission spectra of FITC, NSR-FITC, and NLR-FITC. Arrow denotes FITC embedded into particle.

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研究结果

Fig 3. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) images of three different shaped MSNs. (A, C, E) TEM images of sphere-shaped particles (A), short rod-shaped particles (C) and long rod-shaped particles (E). (B, D, F) SEM images of sphere-shaped particles (B), short rod-shaped particles (D) and long rod-shaped particles (F).

Fig 4. TEM images of cells incubated for 30 min with 50 mg/ml different shaped MSNs. (A) NS100, (B) NSR240, and (C) NLR450 particles. The marked areas are representative images of cross-section views (circled area) and horizontal views (boxed area) of MSNs in cells. (D-F) Illustrations of the pathway for cellular internalization of NS100 (D), NSR240 (E), and NLR450 (F) particles.

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Fig 5. Confocal microscopy images of A375 cells after 4 h incubation at 37 C with NS100 (A–C), NSR240 (D–F) and NLR450 (G–I) particles: Fluorescent images of the cell nucleus (A, D, G), Images of MSN-FITC fluorescence in cells (B, E, H), Image of MSN-FITC fluorescence superimposed on the nucleus (C, F, I).

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Fig 6. Quantification of cellular uptake of different shaped MSNs by fluorescene-activated cell sorting (FACS). (A) Fluorescent intensity of MSNs in cells analyzed by FACS. (B) Quantitative analysis of particle numbers in cells by FACS.

Fig 7. Effect of different shaped MSNs on cellular uptake. Confocal microscopy images of A375 cells after 30 min incubation at 37 C with particles conjugated with FITC or RITC. Cellular uptake of NS100-RITC (A) and NSR240-FITC (B), NSR100-FITC (D) and NLR450-RITC (E), NLR 450-FITC (G) and NSR240-RITC (H) particles. C, F, and I are the merge images of A and B, D and E, G and H, respectively.

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Fig 9. Effect of nonspecific cellular uptake of different shaped MSNs on cytoskeleton formation. Representative images of A375 cells cultured on slides were observed by confocal microscopy. Cells were fluorescently stained for cytoskeletal F-actin fibers (green) after treatment without (A) or with NS100 (B), NSR240 (C) and NLR450 (D) particles.

Fig 8. Effect of nonspecific cellular uptake of different shaped MSNs on cell adhesion. Representative images of cell adhesion on slides were observed after A375 cells were treated without (A) or with (B-D) particles. Numbers of adhesion cells treated with different shaped particles compared with the untreated control.

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Fig 10. Effect of nonspecific cellular uptake of different shaped MSNs on the expression of ICAM-1 and MCAM. Western blot analysis (A) and histogram (C) of the protein expression of ICAM-1 and MCAM in A375 cells untreated or treated with different shaped MSNs. Semiquantitative reverse transcription-PCR analysis (B) and histogram (D) of the mRNA expression of ICAM-1 and MCAM in A375 cells untreated or treated with different shaped MSNs.

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Fig 11. Effect of nonspecific cellular uptake of different shaped MSNs on cell migration. Representative images of migrating cells were observed for 24 h after treatment without (A) or with particles (B–D).

Fig 12. Cytotoxicity of different shaped MSNs on A375 cells. (A) MTT cell proliferation assay showing the effect of varying concentrations of different shaped particles. (B) The effect of varying concentrations of shaped particles on early apoptosis of A375 cells by FACS analysis.

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Fig 13. Biodistribution of different shaped and PEGylated MSN-FITC in liver, spleen, and lung was observed by confocal microscopy images 2 h after intravenous injection. Arrows denote NLRFITC distribution in lung.

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Fig 14. Quantitative analysis of different shaped and PEGylated MSNs in organs and blood by ICP-OES. Relative Si contents in liver, spleen, and kidney at (A) 2 h, (B) 24 h, and (C) 7 d postinjection.

Fig 15. ICP-OES quantitative analysis of the Si content in (A) urine, and (B) feces at 2 h, 24 h, and 7 d after injection.

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Fig 16. Effect of different shaped and PEGylated MSNs on (A,B) hematology and (C,D) serum biochemistry at (A,C) 24 h and (B,D) 18 d. Relative hematology indicators include red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet count (PLT) and white blood cell count (WBC). Related serum biochemistry indicators include alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), alkaline phosphatase (ALP), creatinine (CREA), and blood urea nitrogen (BUN).

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Fig 17. Effect of different shaped and PEGylated MSNs on histopathology of the liver, spleen, lung and kidney at 7 d after intravenous injection.

结论

Tianjin University 2012-04-28

细胞内化率与粒子的形状有关，粒子长径比越大，粒子的内化率越大。

长径比大的粒子对细胞的增殖，凋亡，吸附及迁移的影响更大。

粒子在体内的血液循环时间与形状有关，长径比大的血液循环时间相对较长。

PEG 修饰过的粒子比未修饰的粒子的体内降解速率低，同时 PEG 修饰可以改变粒子在体内的分布。

设计的纳米粒子的生物相容性都比较好。