Distribution of Iron Oxide Core-Titanium Dioxide Shell Nanoparticles in VX2 Tumor Bearing Rabbits Introduced by Two Different Delivery Modalities
<p>Intravenous (IV) and transarterial intra-catheter (IC) injected rabbits’ VX2 tumors and surrounding liver tissue stained by H&E, histochemical (HC) stain (detecting the nanoparticles) and Ki67 Antibodies.</p> "> Figure 2
<p>Details from <a href="#app1-nanomaterials-06-00143" class="html-app">Supplemental Figure S3</a>. (<b>a</b>) IV injected rabbit’s lung showing metastatic nodular tumor deposits disposed in a peripheral location composed of poorly differentiated squamoid cells expanding the alveolar septae (100× H&E); (<b>b</b>) A photomicrograph of the IV injected rabbit lung showing alveolar septae expanded and infiltrated by inflammatory infiltrate, predominantly eosinophils (200× H&E); (<b>c</b>) IC rabbit lung showing marked thickening and destruction of alveolar spetae by metastatic deposits and inflammatory infiltrate composed mainly of granulocytes (arrowheads in inset, 400×). Blood vessels are engorged with granulocytes (arrow head) (100×); (<b>d</b>,<b>e</b>) IV injected rabbit’s splenic red pulp displaying dilated and congested sinusoids. Arrows point to follicles (100× H&E); (<b>f</b>) IC injected rabbit spleen showing congested and expanded red pulp with influx of macrophages and proliferated Littoral cells; (<b>g</b>) IV injected rabbit renal cortex showing mild to moderate cloudy changes in the tubules (100× H&E); (<b>h</b>) IC injected rabbit kidney showing cortical tubules exhibiting severe cloudy swelling and vesicular changes (100× H&E); (<b>i</b>) An IV injected rabbit liver micrograph displaying mild lobular disarray with hepatocyte ballooning (200× H&E); (<b>j</b>) IC injected rabbit’s liver showing dilated congested central veins and sinusoids. Hepatocytes show ballooning and microvesicular steatosis (200× H&E); (<b>k</b>) Photomicrograph showing part of VX2 tumor mass with a central necrotic cavity (100× H&E); (<b>l</b>) A photomicrograph of VX2 tumor showing infiltrating malignant cells on the left side, tumor infiltrating lymphocytes on the right, and a dilated sinusoid with tumor thrombus in the center (arrow) (100× H&E); (<b>m</b>) Photomicrograph of VX2 tumor showing nests of squamous cell carcinoma (yellow arrow), surrounded by a peripheral zone of tumor infiltrating lymphocytes (red arrow), compressed hepatocytes and fibroblasts; (<b>n</b>) Control rabbit lung micrograph showing multiple microscopic metastatic tumor deposits (arrows), congested microvasculature and patent alveoli (100× H&E); (<b>o</b>) Control Rabbit spleen micrograph showing white pulp follicles with narrow intervening red pulp sinusoids, minimal congestion ascribed to manipulation during necropsy; (<b>p</b>) Control rabbit kidney micrograph showing patent cortical tubules and unremarkable glomerular tufts with absence of congested vessels (100× H&E); (<b>q</b>) Photomicrograph of control rabbit liver (100× H&E); (<b>r</b>) Control Rabbit micrograph of VX2 liver tumor showing malignant squamoid cells (arrow) surrounding a central necrotic cavity (star) (100× H&E).</p> "> Figure 3
<p>Details from <a href="#app1-nanomaterials-06-00143" class="html-app">Supplemental Figure S3</a>. Histochemical Staining for NPs: (<b>a</b>) IV injected rabbit’s VX2 tumors showing weak uneven staining of tumor masses. Arrow shows weak staining of surrounding hepatocytes (100×); (<b>b</b>) Photomicrograph showing nuclear staining of IV injected rabbit’s VX2 tumor cells with weak cytoplasmic stain. Surrounding tissues are not stained (200×); (<b>c</b>) Intense brown nuclear and cytoplasmic staining of the IC injected rabbit’s tumor masses (100×); (<b>d</b>) Micrograph of IC injected rabbit’s VX2 tumor showing intense positive nuclear and cytoplasmic staining of cells (400×); (<b>e</b>) Photomicrograph of control rabbit VX2 showing faint non-specific staining of malignant tumor cells (arrow) and necrotic tissues (star); (<b>f</b>) Micrograph of IV injected rabbit’s spleen showing positively stained red pulp macrophages and marginal zone macrophages (arrows) (200×); (<b>g</b>) Micrograph of IC injected rabbit’s spleen showing positively stained red pulp macrophages (400×); (<b>h</b>) Micrograph of control rabbit’s spleen red pulp showing absence of positive staining (200×); (<b>i</b>) Positively stained Kupffer cell in a sinusoid of an IV injected rabbit (400×); (<b>j</b>) IC injected rabbit’s micrographs showing intense staining of periportal hepatocytes and bile duct epithelium (arrow) (200×); (<b>k</b>) Micrograph of control liver showing no positive staining (200×); (<b>l</b>) IV injected rabbit renal medullary tubules showing intense positive cytoplasmic stain (200×); (<b>m</b>) IC injected rabbit’s micrograph of rabbit medullary tubules of kidney showing intense positive cytoplasmic stain (200×); (<b>n</b>) Control rabbit’s kidney micrograph showing no positive staining (100×); (<b>o</b>) IV injected rabbit’s lung showing a nodular metastatic deposit with positively stained tumor cells (200×); (<b>p</b>) IV injected rabbit’s lung displaying positively stained tumor metastatic deposits, in addition to alveolar macrophages. Arrow is pointing at a positively stained alveolar macrophage (200×); (<b>q</b>) IC rabbit lung showing positive brown staining of metastatic deposits within the alveolar septae (red arrowhead) and the bronchiolar epithelial lining cells (black arrowhead) (200×); (<b>r</b>) Micrograph of control rabbit’s lung showing absence of positive staining in metastatic deposits (arrows) and alveolar macrophages (100×).</p> "> Figure 4
<p>(<b>a</b>) NP positive Kupffer cells and (<b>b</b>) total number of Kupffer cells in livers of intravenous (IV) and intra-catheter (IC) injected and control rabbits. <span class="html-italic">Y</span>-axis indicates total number of Kupffer cells per field of view, averaged from 20 such areas, images acquired at 400× (for example, see one such area of view in <a href="#nanomaterials-06-00143-f003" class="html-fig">Figure 3</a>i).</p> "> Figure 5
<p>Overview of XFM images for NP distribution after intravenous (IV) and transarterial intra-catheter (IC) injections into control rabbits VX2 tumor and liver, kidney, lung, and spleen tissues. The mapping for the Titanium (<b>left</b>) and Zinc (<b>right</b>) concentrations and content informs about tissue shape, and potentially even its health status. It should be noted that these are false color images showing distribution of elemental concentrations from the lowest (<b>black to brown</b>) to highest (<b>yellow to white</b>) within the elemental concentrations distribution of each sample (see color scale bar in lower right depicting spectra of colors matching lowest concentration (in a given sample)—black) to highest (<b>white</b>) across a “red temperature” scale. Thus, samples with very low elemental concentrations for a given element (e.g., Ti in control samples) show a “salt and pepper” pixel distribution indicating that background signal levels predominate in the sample. It should be noted that paraffin embedding disrupts distribution of free ions such as potassium, while sulfur and zinc distributions still well represent a tissue outline because Zn finger proteins are accumulated in each cells’ nucleus [<a href="#B14-nanomaterials-06-00143" class="html-bibr">14</a>,<a href="#B15-nanomaterials-06-00143" class="html-bibr">15</a>,<a href="#B16-nanomaterials-06-00143" class="html-bibr">16</a>,<a href="#B17-nanomaterials-06-00143" class="html-bibr">17</a>,<a href="#B18-nanomaterials-06-00143" class="html-bibr">18</a>,<a href="#B19-nanomaterials-06-00143" class="html-bibr">19</a>]. Distribution of nanoparticles, much larger and insoluble <span class="html-italic">in situ</span>, is considered to be accurately depicted by XFM [<a href="#B14-nanomaterials-06-00143" class="html-bibr">14</a>,<a href="#B15-nanomaterials-06-00143" class="html-bibr">15</a>,<a href="#B16-nanomaterials-06-00143" class="html-bibr">16</a>,<a href="#B17-nanomaterials-06-00143" class="html-bibr">17</a>,<a href="#B18-nanomaterials-06-00143" class="html-bibr">18</a>,<a href="#B19-nanomaterials-06-00143" class="html-bibr">19</a>].</p> "> Figure 6
<p>Graphic representation of data in <a href="#nanomaterials-06-00143-t001" class="html-table">Table 1</a>. (<b>Top</b>) Ti concentration (in femtograms per pixel) in different tissues from nanoparticle treated animals and controls; data for each animal are shown. (<b>Bottom</b>) Mean ratio of Ti (femtograms) vs. S (femtograms) per pixel; please note that data structure allows comparison on a per pixel basis. Thus, information provided by the bottom graph allows us to monitor the situation most accurately, e.g., if tissue integrity is uneven, resultant uneven concentration of nanoparticles in tissue will not be misinterpreted. Likewise, false background (“salt and pepper” signal pattern for Ti) is effectively removed when false signals are divided by real signal for S.</p> ">
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Animal Model and Tumor Induction
4.2. Nanoparticle Preparation
4.3. Nanoparticle Delivery
4.4. Tissue Sample Preparation
4.5. Histochemical Detection of Nanoparticle Distribution
4.6. X-ray Fluorescence Microscopy (XFM)
4.7. Inductively Coupled Plasma Mass Spectrometry (ICP-MS)
4.8. Immunohistochemistry
4.9. Semi-Quantitative Analysis of Ki67 and TUNEL Indices
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
ICP-MS | Inductively coupled plasma mass spectrometry |
XFM | X-ray fluorescence microscopy |
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Elemental Content Ratio | Mean | SD | ||||
---|---|---|---|---|---|---|
IC | IV | p | IC | IV | ||
VX2 | Ti/P * | 0.01711 | 0.02127 | 0.26872 | 0.00297 | 0.00476 |
Ti/S | 0.01528 | 0.01383 | 0.84901 | 0.00224 | 0.01217 | |
Ti/Zn | 0.64011 | 0.65397 | 0.89887 | 0.14092 | 0.10773 | |
Ti/Fe | 0.12339 | 0.1504 | 0.27581 | 0.02431 | 0.02803 | |
Liver | Ti/P | 0.01262 | 0.02182 | 0.03075 *** | 0.00373 | 0.00313 |
Ti/S | 0.00736 | 0.0137 | 0.00965 | 0.0023 | 0.00052 | |
Ti/Zn | 0.26631 | 0.45213 | 0.02825 | 0.08716 | 0.03959 | |
Ti/Fe | 0.06452 | 0.09442 | 0.0991 | 0.02088 | 0.01223 | |
Spleen | Ti/P | 0.0149 | 0.03427 | 0.0403 | 0.00564 | 0.00969 |
Ti/S | 0.01895 | 0.04221 | 0.04122 | 0.00697 | 0.01165 | |
Ti/Zn | 0.85428 | 2.125 | 0.03773 | 0.50079 | 0.51691 | |
Ti/Fe | 0.04296 | 0.08506 | 0.13063 | 0.0064 | 0.0379 | |
Lung | Ti/P | 0.02079 | 0.02239 | 0.8848 | 0.01005 | 0.01489 |
Ti/S | 0.00803 | 0.01081 | 0.28208 | 0.00107 | 0.00373 | |
Ti/Zn | 0.44638 | 0.61228 | 0.4479 | 0.1362 | 0.31355 | |
Ti/Fe | 0.07881 | 0.08918 | 0.77821 | 0.04038 | 0.04388 | |
Kidney | Ti/P | 0.01028 | 0.0146 | 0.11988 | 0.00219 | 0.0031 |
Ti/S | 0.00568 | 0.0087 | 0.10969 | 0.0018 | 0.0018 | |
Ti/Zn | 0.20283 | 0.40091 | 0.03195 | 0.05167 | 0.09277 | |
Ti/Fe | 0.07926 | 0.13716 | 0.06617 | 0.02248 | 0.03306 | |
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Refaat, T.; West, D.; El Achy, S.; Parimi, V.; May, J.; Xin, L.; Harris, K.R.; Liu, W.; Wanzer, M.B.; Finney, L.; et al. Distribution of Iron Oxide Core-Titanium Dioxide Shell Nanoparticles in VX2 Tumor Bearing Rabbits Introduced by Two Different Delivery Modalities. Nanomaterials 2016, 6, 143. https://doi.org/10.3390/nano6080143
Refaat T, West D, El Achy S, Parimi V, May J, Xin L, Harris KR, Liu W, Wanzer MB, Finney L, et al. Distribution of Iron Oxide Core-Titanium Dioxide Shell Nanoparticles in VX2 Tumor Bearing Rabbits Introduced by Two Different Delivery Modalities. Nanomaterials. 2016; 6(8):143. https://doi.org/10.3390/nano6080143
Chicago/Turabian StyleRefaat, Tamer, Derek West, Samar El Achy, Vamsi Parimi, Jasmine May, Lun Xin, Kathleen R. Harris, William Liu, Michael Beau Wanzer, Lydia Finney, and et al. 2016. "Distribution of Iron Oxide Core-Titanium Dioxide Shell Nanoparticles in VX2 Tumor Bearing Rabbits Introduced by Two Different Delivery Modalities" Nanomaterials 6, no. 8: 143. https://doi.org/10.3390/nano6080143