Name: establishment of a simple and effective rat model for intraoperative parathyroid gland imaging
Uploaded: 2022-08-17T14:05:00
Description: Watch this Scientific Journal Video about Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging at JoVE.com

This study was supported by the National Natural Science Foundation of China (NSFC) (82172598), the Natural Science Foundation of Zhejiang Province, China (LZ22H310001), the 551 Health Talent Training Project of Health Commission of Zhejiang Province, China, and the Medical and Health Science and Technology Project of Zhejiang Province, China (2021KY110).

All animal studies have been approved by the Institutional Animal Care and Use Committee (IACUC) of the Institute of Basic Medicine and Cancer, Chinese Academy of Sciences. This is a non-survival surgery.
1. Animal
<ol>
	<li>Use a 6-8-week-old female SD rat, weighing 250 g, for intraoperative PG imaging.</li>
</ol>
2. Anesthesia
<ol>
	<li>Turn on the anesthesia machine.</li>
	<li>Before beginning, ensure the isoflur...

<ol>
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H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidental+parathyroidectomy+during+thyroid+surgery:+an+underappreciated+complication+of+thyroidectomy.">Incidental parathyroidectomy during thyroid surgery: an underappreciated complication of thyroidectomy.</a> World Journal of Surgery. 29 (12), 1539-1543 (2005).</li><li>Sahyouni, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rate+of+incidental+parathyroidectomy+in+a+pediatric+population.">Rate of incidental parathyroidectomy in a pediatric population.</a> OTO Open. 5 (4), (2021).</li><li>Erickson, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence,+survival+time,+and+surgical+treatment+of+parathyroid+carcinomas+in+dogs:+100+cases+(2010-2019).">Incidence, survival time, and surgical treatment of parathyroid carcinomas in dogs: 100 cases (2010-2019).</a> Journal of the American Veterinary Medical Association. 259 (11), 1309-1317 (2021).</li><li>Bi, R., Fan, Y., Luo, E., Yuan, Q., Mannstadt, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+techniques+to+create+hypoparathyroid+mice:+parathyroidectomy+using+GFP+glands+and+diphtheria-toxin-mediated+parathyroid+ablation.">Two techniques to create hypoparathyroid mice: parathyroidectomy using GFP glands and diphtheria-toxin-mediated parathyroid ablation.</a> Journal of Visualized Experiments. (121), e55010 (2017).</li><li>Soulsby, S. 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We demonstrate an IONPs-guided negative imaging technique of rat PG&#160;using black IONPs, which were injected locally in the center of the thyroid gland and diffused within the thyroid gland but not the PG. It enables clear identification of the PG&#160;with naked eyes without the aid of any microscope. Although transgenic mice with green fluorescent protein selectively expressed in the PG&#160;have been reported13, the model described in this paper is more straightforward to perform. It only ta...

Parathyroid gland&#160;(PG) is small, oval-shaped endocrine glands located in the neck of humans and other vertebrates, which produce and secrete parathyroid hormones to regulate and balance calcium and phosphorus levels in the blood and in bones1. Humans usually have two pairs of PG&#160;located behind the thyroid gland lobes in variable locations; the size of human PG&#160;typically measures 6 mm x 4 mm x 2 mm, with a weight of approximately 35-40 mg2. Removal or damage of the PG&#160;causes hypoparathyroidism (HP), an endocrine disorder characterized by hypocalcemia and low or undetectable levels of parathyroid hormones, which cause a wide range of symptoms from cramp-like spasms to malformed teeth to chronic kidney diseases. Some of these complications are fatal (e.g., heart failure and seizure)3,4,5; thus, PG&#160;is essential in regulating the body&#39;s metabolism and sustaining life.
HP is one of the most common complications after anterior neck surgery, especially in thyroidectomy, a well-established curative treatment for thyroid cancer, which is the most common endocrine cancer worldwide6,7. Post-thyroidectomy HP is predominantly caused by direct trauma, ischemia, or removal of the PG in surgery because of a severe lack of ability to reliably discriminate the PG&#160;from thyroid gland lobes and other surrounding tissues (e.g., lymph nodes and peripheral fat particles) in real-time in the operation room. In 2021, Barrios et al. reported an average PG mis-section rate of 22.4% within 1,114 thyroidectomy cases, and even experienced surgeons who had a minimum error rate of 7.7%8. Such high PG mis-section rates are consistent with other similar reports9,10,11. Thus, incorrect parathyroidectomy is an independent risk factor for transient and permanent postoperative HP.
Developing effective intraoperative PG identification methods holds the key to addressing this critical unmet medical need; however, it has been severely limited by the lack of animal models in preclinical research. To date, most intraoperative PG identification investigations have been performed on human patients and large animals (e.g., dogs)12, which are expensive and difficult to receive ethical approval, expand subject numbers, and repeat tests. Meanwhile, the mouse, the most commonly used vertebrate model in biological research, has extremely small PG, with a size of less than 1 mm13. Due to this limitation, mouse PG models have seldom been used in intraoperative PG identification research.
This paper reports the establishment of a simple, straightforward, and effective rat model for intraoperative PG identification studies. We investigated the usage of native Sprague-Dawley (SD) rats without any surgical modifications or genetic engineering as a reliable animal model for testing a PG imaging contrast agent, IONPs, in a thyroidectomy surgery. This rat model demonstrates a highly similar physiological structure of PG&#160;and the surrounding microenvironment to that of humans, and the size of rat PG&#160;is large enough to be visually detected in comparison with those of mice. Most rats have one PG on each side of the thyroid gland. The simplicity and effectiveness of this rat model have been demonstrated by performing intraoperative IONP-enhanced PG imaging in thyroidectomy surgery.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
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			<td>alcohol</td>
			<td>Li feng</td>
			<td>9400820067</td>
			<td></td>
		</tr>
		<tr>
			<td>anesthesia machine</td>
			<td>RWD Company</td>
			<td>R520IE</td>
			<td>Machine number</td>
		</tr>
		<tr>
			<td>blade</td>
			<td>Daopian</td>
			<td>TB-JZ-10#</td>
			<td></td>
		</tr>
		<tr>
			<td>cylindrical pillow</td>
			<td></td>
			<td></td>
			<td>made by ourselves</td>
		</tr>
		<tr>
			<td>depilatory cream</td>
			<td>Nair</td>
			<td>TMG-001</td>
			<td></td>
		</tr>
		<tr>
			<td>electronic scale</td>
			<td>Hong xingda</td>
			<td>CN-HXD2</td>
			<td></td>
		</tr>
		<tr>
			<td>eosin</td>
			<td>Thermo Fisher (Waltham, USA).</td>
			<td>C0105S-2</td>
			<td></td>
		</tr>
		<tr>
			<td>erythromycin</td>
			<td>Shuang ji (Beijing, China)</td>
			<td>200409</td>
			<td></td>
		</tr>
		<tr>
			<td>gauze</td>
			<td>Fulanns</td>
			<td>YY0331-2006</td>
			<td></td>
		</tr>
		<tr>
			<td>heating pad</td>
			<td>Johon (ShenZhen,China)</td>
			<td>JH-36-2006</td>
			<td></td>
		</tr>
		<tr>
			<td>hematoxylin</td>
			<td>Thermo Fisher (Waltham, USA).</td>
			<td>C0105S-1</td>
			<td></td>
		</tr>
		<tr>
			<td>insulin injection needle</td>
			<td>Jiangyin NanquanMacromolecule</td>
			<td>20170702</td>
			<td></td>
		</tr>
		<tr>
			<td>iodophor cotton ball</td>
			<td>HOYON</td>
			<td>19-6007</td>
			<td></td>
		</tr>
		<tr>
			<td>iron oxide nanoparticle solution</td>
			<td>Zhongke Leiming Technology (Beijing, China)</td>
			<td>Mag9110-05</td>
			<td></td>
		</tr>
		<tr>
			<td>isoflurane</td>
			<td>Sigma Aldrich (St Louis USA).</td>
			<td>21112801</td>
			<td></td>
		</tr>
		<tr>
			<td>needle holder</td>
			<td>Meijun</td>
			<td>MH0587</td>
			<td></td>
		</tr>
		<tr>
			<td>operation table</td>
			<td>BioJane</td>
			<td>BJ-P-M</td>
			<td></td>
		</tr>
		<tr>
			<td>paraformaldehyde solution</td>
			<td>Biosharp</td>
			<td>21269333</td>
			<td></td>
		</tr>
		<tr>
			<td>rubber</td>
			<td>G-CLONE</td>
			<td> 
			XT41050</td>
			<td></td>
		</tr>
		<tr>
			<td>scanning machine</td>
			<td>Olympus</td>
			<td>Slideview VS200</td>
			<td></td>
		</tr>
		<tr>
			<td>surgical forceps</td>
			<td>Suping</td>
			<td>SPHC-0676</td>
			<td></td>
		</tr>
		<tr>
			<td>surgical knife handle</td>
			<td>Aladdin</td>
			<td>S3052-06-1EA</td>
			<td></td>
		</tr>
		<tr>
			<td>surgical retractor</td>
			<td>TOCYTO</td>
			<td>18-4010</td>
			<td></td>
		</tr>
		<tr>
			<td>surgical scissors</td>
			<td>Suping</td>
			<td>SPHC-0795</td>
			<td></td>
		</tr>
		<tr>
			<td>surgical towel</td>
			<td>Along technology</td>
			<td>YCKJ-RJ-036205</td>
			<td></td>
		</tr>
		<tr>
			<td>suture</td>
			<td>Ethicon</td>
			<td>SA84G</td>
			<td></td>
		</tr>
		<tr>
			<td>suture with needle</td>
			<td>Jinhuan (Shanghai,China)</td>
			<td>F301</td>
			<td></td>
		</tr>
		<tr>
			<td>vascular forceps</td>
			<td>Along technology</td>
			<td>YCKJ-RJ-016218</td>
			<td></td>
		</tr>
		<tr>
			<td>Water Bath-Slide Drier</td>
			<td>Hua su (Jinhua, China)</td>
			<td>HS-1145</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishment of a simple and effective rat model for intraoperative parathyroid gland imaging

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is similar in color to the thyroid gland. The lack of effective animal models in preclinical research is a severe limitation for the development of PG identification techniques. This protocol allows for the establishment of a simple and effective rat model for PG identification. In this model, black iron oxide nanoparticles (IONPs) are injected locally in the thyroid gland and rapidly diffuse within the thyroid gland but not the PG. A negatively stained PG and a positively stained thyroid gland can be easily identified by the naked eye without requiring external microscopes. The position of the PG can be identified by increasing the color contrast between the thyroid gland and the PG, based on the color of the black IONPs. This rat model is low-cost and convenient for PG identification, and the IONPs are a novel PG contrast agent.

In this animal model, we surgically incised the neck of a SD rat to expose the trachea, larynx, and surrounding tissues. Then, the thyroid gland was visually located on both sides of the trachea; it is butterfly-shaped and approximately 3 mm x 5 mm in size. A pair of PG&#160;are usually located in the upper part of the thyroid gland, and their color is very similar to that of the thyroid gland lobes, making it extremely difficult to distinguish them with the naked eye (Figure 1).&#160;
<...

Watch this Scientific Journal Video about Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging at JoVE.com

Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging

To date, the development of parathyroid gland (PG) identification methods is limited by the lack of animal models in preclinical research. Here, we establish a simple and effective rat model for intraoperative PG imaging and evaluate its effectiveness by using iron oxide nanoparticles as a novel PG contrast agent.

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is ...

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