The present study and production to this article were funded by the National Natural Science Foundation of China (Grant 81773373, 81172614 and Grant 81441089).&#160;We thank Dr. Jin Yan&#160;and Dr. Pan Yujie, of Department of Emergency, Beijing Chaoyang Hospital,&#160;and Dr. Qu Peng of Department of Ultrasound Medicine, Beijing Chaoyang Hospital&#160;for&#160;helping with the video production.&#160;

The study followed guidelines of Capital Medical University (Beijing, P.R China) for the care and use of experimental animals. All procedures were approved by the Animal Ethical Committee of Capital Medical University in China.
1. Experimental preparations
NOTE: Acclimate the female specific pathogen-free Wistar rats (weight: 200 &#177; 10 g) to the experimental environments for a week before administration (Environmental conditions: light/dark:12h/12h, temperature 22...

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The authors have nothing to disclose.

Sonography is the most convenient tool for determining pulmonary diseases, due to its excellent sensitivity to the free fluid in the pleural cavity11. That is because sonography can immediately detect the contrast in acoustic impedance of air and fluids in the lung12. Besides, sonography is more flexible in a small animal&#39;s model than CT. Nevertheless, the air in the lung reflected the sound wave and impeded from observing the intrapulmonary changes after nanoparticles ...

Pleural effusion is a very common clinical manifestation of pulmonary diseases with a variety of causes. Having a useful animal pleural effusion model is very important to study these pulmonary diseases, the roles of the two pleural membrane layers, the mechanisms of pleural effusion, and its treatment. However, some reported pleural effusion models mainly focus on the malignant pleural effusion or biological factors rather than the nanoparticles in the environment1,2. Here, we introduce a new model of pleural effusion that is simple, safe and effective.
With the development of nanotechnology and the extensive use of nanoproducts, there is a concern about the potential hazards of nanomaterials to the environment and the human health3,4. Nanomaterials introduce risk factors and potentially lead to novel hazards within the workplace or through environmental contamination. In vitro and in vivo studies show that nanomaterials can result in multi-organ damage to the lungs, the heart, the liver, the kidney, and the nervous system, as well as the reproductive and immune systems5,6. Additionally, some studies reported that the specific toxicity of nanomaterials was due to their unique physicochemical properties3,4,7.
We have reported that a group of workers with occupational exposure to nanomaterials clinically presented with pleural and pericardial effusion, pulmonary fibrosis and granuloma8,9. Silica nanoparticles were isolated in these patients&#39; pleural effusion9. In order to reproduce and verify the pleural effusion induced by the inhaled nanoparticles in human, we conducted the experiment by instilling the polyacrylate/nanosilica (PA/NPSi) via the respiratory tract in rats, which mimicked human respiration in a real environment, and found that intratracheal instillation of PA/NPSi could result in pleural effusion in rats. Here, we introduce how to make pleural effusion in rats by intratracheal instillation of PA/NPSi, and how to isolate nanoparticles in the pleural effusion. This model may be useful for the further study of nanotoxicology and pleural effusion diseases.

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	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:227px;">Acuson S2000 Color Doppler ultrasound system</td>
			<td style="width:196px;">Siemens Medical Solutions, Mountain View ,CA</td>
			<td style="width:160px;"></td>
			<td style="width:260px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:227px;">&#160;Polyacrylate/nanosilica</td>
			<td style="width:196px;">Fudan University,Shanghai, China</td>
			<td style="width:160px;"></td>
			<td>made by order with nanosilica(20&#177;5)nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10% chloral hydrate</td>
			<td style="width:196px;">Beijing Chemical Works</td>
			<td style="width:160px;">302-17-0</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:227px;">Transmission electron microscope&#160;</td>
			<td style="width:196px;">JEM-1400Plus&#65292;JEOL Ltd., Japan.</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:227px;">Light speed 16 spiral computed tomography</td>
			<td style="width:196px;">GE Healthcare, US</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:227px;">Specific pathogen-free Wistar</td>
			<td style="width:196px;">Animal Center of Lianhelihua (Beijing, China)</td>
			<td style="width:160px;"></td>
			<td>Wistar rats</td>
		</tr>
	</tbody>
</table>

a pleural effusion model in rats by intratracheal instillation of polyacrylate/nanosilica

Pleural effusion is a prevalent clinical finding of many pulmonary diseases. Having a useful animal pleural effusion model is very important to study these pulmonary diseases. Previous pleural effusion models paid more attention to biological factors rather than nanoparticles in the environment. Here, we introduce a model to make pleural effusion in rats by intratracheal instillation of polyacrylate/nanosilica, and a method of nanoparticle isolation in the pleural effusion. By intratracheal instillation of polyacrylate/nanosilica with concentrations of 3.125, 6.25 and 12.5 mg/kg&#8729;mL, the pleural effusion in rats presented on day 3, peaked at days 7-10 in 6.25 and 12.5 mg/kg&#8729;mL groups, then slowly decreased and disappeared on day 14. When the concentration of polyacrylate/nanosilica increased, the pleural effusion is produced more and faster. This pleural fluid was detected by ultrasound examination or CT chest scanning and confirmed by dissection of rats. Silica nanoparticles were observed in the rats' pleural effusion by transmission electron microscope. These results showed that the exposure to polyacrylate/nanosilica leads to the induction of pleural effusion, which was consistent with our previous report in humans. Additionally, this model is beneficial for the further study of nanotoxicology and the pleural effusion diseases.

Using a thoracic ultrasound, we found no pleural effusions on day 1 in all groups. However, on day 3, the pleural effusion appeared in the 6.25 and 12.5 mg/kg&#8729;mL groups. The effusion was mainly in the right costal phrenic angle, while the pericardial effusion only presented in 12.5 mg/kg&#8729;mL group. Furthermore, on day 7, both pleural effusion (Video 1) and pericardial effusion (Video 2) were detected in 6.25 mg/kg&#8729;mL group (<strong class=...

Watch this Scientific Journal Video about A Pleural Effusion Model in Rats by Intratracheal Instillation of Polyacrylate/Nanosilica at JoVE.com

A Pleural Effusion Model in Rats by Intratracheal Instillation of Polyacrylate/Nanosilica

Here, we present a protocol to construct a pleural effusion model in rats by intratracheal instillation of polyacrylate/nanosilica.

Pleural effusion is a prevalent clinical finding of many pulmonary diseases. Having a useful animal pleural effusion model is very important to study ...

This method can provide insight into the toxicity of nanomaterials and can be applied for the study of pulmonary toxicity. This technique was derived to study the effects of polyacrylate/nanosilica-induced pleural effusion. Before beginning the instillation, sonicate a fresh, 10-milliliter, polyacrylate/nanosilica suspension in normal saline for 20 to 30 minutes, followed by 10 minutes of vortexing, before diluting the suspension to 3.125, 6.25, and 12.5 milligram per milliliter concentrations.Next, confirm a lack of response to toe pinch in an anesthetized rat, and fix the front teeth of the animal to the board with a piece of nylon string. Using forceps and a frontal lets, gently open the mouth to visualize the fissure of glottis, and insert a fine tube into the bilateral bronchus. Then, instill 0.5 milliliters of the polyacrylate/nanosilica particles through the tube into the lungs, and place the rat in the supine position with monitoring until full recovery.On days one, three, seven, and 14 after instillation, use an electric shaver to remove the hair from the chest and upper abdomen of each anesthetized rat, and place the rat on the mounting plate of an ultrasound system with a linear array transducer in the supine position. Apply ultrasound gel to the exposed skin, and place the transducer on the intercostal space and subcostal area to detect the pleural fluid. In order to detect the effusion accurately, select the left and right lateral positions to perform an ultrasound examination.On day seven, both pleural effusion and pericardial effusion are detected in the 6.25 milligram per kilogram per milliliter instilled group. On day seven and 14, CT chest scans of the 12.5 milligram per kilogram per milliliter instilled animals are abnormal at the blunt posterior costophrenic angle, hinting at a small amount of pleural effusion. Transmission electron microscopy of pleural effusion samples reveals nanosilica nanoparticles individually and in clusters with a morphology consistent with that observed for nanoparticles within the prepared suspension.The most important thing is to remember to visualize the fissure of the glottis and to insert a fine tube into the bilateral bronchus. The nanoparticles can also be instilled by inhalation or intravenous injection, as both methods have been performed in previous studies. This technique is useful for studying the development of pleural effusion.

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7.8K 조회수.  Capital Medical University. 이 방법은 나노 물질의 독성에 대한 통찰력을 제공 할 수 있으며 폐 독성연구에 적용 될 수있다. 이 기술은 폴리아크릴레이트/나노실리카 유도 흉막 유출의 효과를 연구하기 위해 유래되었다. 주입을 시작하기 전에, 20~30분 동안 정상 식염수에 10밀리리터, 폴리아크릴레이트/나노실리카 서스펜션을 초음파 처리한 다음 10분간 의소용돌이를 한 후 서스펜션을 3.125, 6.25, 밀리리?...