Cytotoxicity, Uptake Behaviors, and Oral Absorption of Food Grade Calcium Carbonate Nanomaterials
"> Figure 1
<p>Scanning electron microscopy (SEM) images, atomic force microscope (AFM) images, height profiles, and 3D images of (<b>A</b>) food bulk CaCO<sub>3</sub>; (<b>B</b>) food nano CaCO<sub>3</sub>; and (<b>C</b>) SS CaCO<sub>3</sub>.</p> "> Figure 2
<p>Hydrodynamic diameter of food bulk CaCO<sub>3</sub> (dashed line), food nano CaCO<sub>3</sub> (solid line) and SS CaCO<sub>3</sub> (dotted line) as a function of differntial intensity. Horizontal line stands for the position of full-width at half-maximum to evaluate peak broadness.</p> "> Figure 3
<p>Effect of three different types of CaCO<sub>3</sub> particles on cell proliferation of human intestinal INT-407 cells, as measured by water-soluble tetrazolium salts (WST-1) assay. (<b>A</b>) Cell proliferation exposed to 250 μg/mL particles or an equivalent amount of CaCl<sub>2</sub> (based on calcium content) for 1–24 h; (<b>B</b>) Cell proliferation treated with different concentrations of CaCO<sub>3</sub> particles or CaCl<sub>2</sub> for 24 h.</p> "> Figure 4
<p>Effect of three different types of CaCO<sub>3</sub> particles or an equivalent amount of CaCl<sub>2</sub> (based on calcium content) on (<b>A</b>) ROS generation and (<b>B</b>) lactate dehydrogenase (LDH) release from human intestinal INT-407 cells after 24 h of incubation. The mean values with different letters (a, a,b, b, c) at the same concentration or time points indicate statistically significant difference (<span class="html-italic">p</span> < 0.05).</p> "> Figure 5
<p>(<b>A</b>) Cellular internalization of three different types of CaCO<sub>3</sub> particles or an equivalent amount of CaCl<sub>2</sub> (based on calcium content) in human intestinal INT-407 cells after 2 h of incubation, as measured by inductively coupled plasma-atomic emission spectroscopy (ICP-AES); (<b>B</b>) intracellular Ca<sup>2+</sup> levels monitored with Calcium Green™<sup>−1</sup> probe (Life Technologies, Carsbad, CA, USA). The mean values with different letters (a, a,b, b) at the same temperature or time points indicate statistically significant difference (<span class="html-italic">p</span> < 0.05). * denotes significant difference in uptake amount between 37 and 4 °C (<span class="html-italic">p</span> < 0.05).</p> "> Figure 6
<p>Intestinal transport of three different types of CaCO<sub>3</sub> particles or an equivalent amount of CaCl<sub>2</sub> (based on calcium content) by microfold (M) cells using an <span class="html-italic">in vitro</span> model of human FAE after 6 h of incubation, as measured by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The mean values with different letters (a, a,b, b) in tested groups indicate statistically significant difference (<span class="html-italic">p</span> < 0.05).</p> "> Figure 7
<p>Plasma concentration-time curves of three different types of CaCO<sub>3</sub> particles (250 μg/mL) or an equivalent amount of CaCl<sub>2</sub> (based on calcium content) after a single-dose oral administration to female rats. Biokinetic data are presented as increase in calcium levels after subtracting the basal plasma calcium levels detected in untreated controls.</p> ">
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization
2.2. Cytotoxicity
2.2.1. Cell Proliferation
2.2.2. Reactive oxygen species (ROS) Generation and lactate dehydrogenase (LDH) Release
2.3. Cellular Uptake Behaviors
2.3.1. Cellular Uptake
2.3.2. Intestinal Transport
2.4. Biokinetics
Biokinetic parameters | Food bulk CaCO3 | Food nano CaCO3 | SS CaCO3 | CaCl2 |
---|---|---|---|---|
Cmax (μg/mL) | 13.39 ± 1.63 c | 21.55 ± 6.71 b,c | 31.56 ± 0.99 b | 66.16 ± 12.98 a |
Tmax (h) | 2.00 c | 1.00 b | 1.00 b | 0.25 a |
AUC (h × μg/mL) | 63.21 ± 2.04 b | 62.26 ± 2.08 b | 66.40 ± 8.60 b | 120.98 ± 11.14 a |
T1/2 (h) | 2.50 ± 0.01 c | 2.86 ± 0.22 d | 1.59 ± 0.01 b | 0.97 ± 0.07 a |
MRT (h) | 4.58 ± 0.18 c | 4.28 ± 0.23 b | 2.94 ± 0.07 a | 2.81 ± 0.20 a |
Absorption (%) 1 | 4.86 ± 0.16 b | 4.79 ± 0.16 b | 5.11 ± 0.66 b | 8.07 ± 0.74 a |
3. Experimental Section
3.1. Materials and Characterization
3.2. Cell Culture
3.3. Cell Proliferation
3.4. Intracellular ROS Generation
3.5. LDH Leakage
3.6. Cellular Uptake
3.7. Intestinal Transport Mechanism
3.8. Oral Absorption
3.9. Statistical Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Kim, M.-K.; Lee, J.-A.; Jo, M.-R.; Kim, M.-K.; Kim, H.-M.; Oh, J.-M.; Song, N.W.; Choi, S.-J. Cytotoxicity, Uptake Behaviors, and Oral Absorption of Food Grade Calcium Carbonate Nanomaterials. Nanomaterials 2015, 5, 1938-1954. https://doi.org/10.3390/nano5041938
Kim M-K, Lee J-A, Jo M-R, Kim M-K, Kim H-M, Oh J-M, Song NW, Choi S-J. Cytotoxicity, Uptake Behaviors, and Oral Absorption of Food Grade Calcium Carbonate Nanomaterials. Nanomaterials. 2015; 5(4):1938-1954. https://doi.org/10.3390/nano5041938
Chicago/Turabian StyleKim, Mi-Kyung, Jeong-A. Lee, Mi-Rae Jo, Min-Kyu Kim, Hyoung-Mi Kim, Jae-Min Oh, Nam Woong Song, and Soo-Jin Choi. 2015. "Cytotoxicity, Uptake Behaviors, and Oral Absorption of Food Grade Calcium Carbonate Nanomaterials" Nanomaterials 5, no. 4: 1938-1954. https://doi.org/10.3390/nano5041938