Name: a silver nanoparticle method for ameliorating biliary atresia syndrome in mice
Uploaded: 2018-10-13T13:30:00
Description: Watch this Scientific Journal Video about A Silver Nanoparticle Method for Ameliorating Biliary Atresia Syndrome in Mice at JoVE.com

The AgNPs used here were a gift from C. M. Che in the Department of Chemistry, the University of Hong Kong. This work was funded by the National Natural Science Foundation of China (No. 81600399) and the Science and Technology Project of Guangzhou (No.201707010014).

All animal experimental protocols have been approved by the Institutional Animal Care and Use Committee of the Sun Yat-Sen University Laboratory Animal Center (#IACUC-DB-16-0602).
1. Establishing the Biliary Atresia Mouse Model
<ol>
	<li>Maintain pregnant BALB/c mice in a specific pathogen-free environment under a 12 h dark/light cycle at 25 &#176;C, with access to autoclaved chow ad libitum.</li>
	<li>To prepare the RRV strain MMU 18006, amplify the virus in MA104 cells and measure...

<ol>
	<li>Szavay, P. O., Leonhardt, J., Czechschmidt, G., Petersen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+reovirus+type+3+infection+in+an+established+murine+model+for+biliary+atresia.">The role of reovirus type 3 infection in an established murine model for biliary atresia.</a> European Journal of Pediatric Surgery. 12 (04), 248-250 (2002).</li><li>Coots, A., 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=Rotavirus+infection+of+human+cholangiocytes+parallels+the+murine+model+of+biliary+atresia.">Rotavirus infection of human cholangiocytes parallels the murine model of biliary atresia.</a> Journal of Surgical Research. 177 (2), 275-281 (2012).</li><li>Shanmugam, N. P., Jayanthi, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biliary+atresia+with+cytomegalovirus.">Biliary atresia with cytomegalovirus.</a> Indian Pediatrics. 49 (2), 157 (2012).</li><li>Mack, C. L., Feldman, A. G., Sokol, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clues+to+the+Etiology+of+Bile+Duct+Injuryin+Biliary+Atresia.">Clues to the Etiology of Bile Duct Injuryin Biliary Atresia.</a> Seminars in Liver Disease. 32, 307-316 (2012).</li><li>Muraji, T., Suskind, D. L., Irie, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biliary+atresia:+a+new+immunological+insight+into+etiopathogenesis.">Biliary atresia: a new immunological insight into etiopathogenesis.</a> Expert Review of Gastroenterology &amp; Hepatology. 3 (6), 599-606 (2009).</li><li>Sokol, R. J., Mack, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiopathogenesis+of+Biliary+Artesia.">Etiopathogenesis of Biliary Artesia.</a> Seminars in Liver Disease. 21, 517-524 (2001).</li><li>Shivakumar, P., Sabla, G. E., Whitington, P., Chougnet, C. A., Bezerra, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+NK+cells+target+the+mouse+duct+epithelium+via+Nkg2d+and+drive+tissue-specific+injury+in+experimental+biliary+atresia.">Neonatal NK cells target the mouse duct epithelium via Nkg2d and drive tissue-specific injury in experimental biliary atresia.</a> Journal of Clinical Investigation. 119 (8), 2281-2290 (2009).</li><li>Mack, C. L., 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=Oligoclonal+expansions+of+CD4++and+CD8++T-cells+in+the+target+organ+of+patients+with+biliary+atresia.">Oligoclonal expansions of CD4+ and CD8+ T-cells in the target organ of patients with biliary atresia.</a> Gastroenterology. 133 (1), 278-287 (2007).</li><li>Shivakumar, P., 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=Effector+Role+of+Neonatal+Hepatic+CD8+++Lymphocytes+in+Epithelial+Injury+and+Autoimmunity+in+Experimental+Biliary+Atresia.">Effector Role of Neonatal Hepatic CD8 + Lymphocytes in Epithelial Injury and Autoimmunity in Experimental Biliary Atresia.</a> Gastroenterology. 133 (1), 268-277 (2007).</li><li>Saxena, V., 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=Dendritic+Cells+Regulate+Natural+Killer+Cell+Activation+and+Epithelial+Injury+in+Experimental+Biliary+Atresia.">Dendritic Cells Regulate Natural Killer Cell Activation and Epithelial Injury in Experimental Biliary Atresia.</a> Science Translational Medicine. 3 (102), (2011).</li><li>Miethke, A. 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=Post-natal+paucity+of+regulatory+T+cells+and+control+of+NK+cell+activation+in+experimental+biliary+atresia.">Post-natal paucity of regulatory T cells and control of NK cell activation in experimental biliary atresia.</a> Journal of Hepatology. 52 (5), 718-726 (2010).</li><li>Tian, J., 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=Topical+delivery+of+silver+nanoparticles+promotes+wound+healing.">Topical delivery of silver nanoparticles promotes wound healing.</a> ChemMedChem. 2 (1), 129-136 (2010).</li><li>Lu, L., 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=Silver+nanoparticles+inhibit+hepatitis+B+virus+replication.">Silver nanoparticles inhibit hepatitis B virus replication.</a> Antiviral Therapy. 13 (2), 253-262 (2008).</li><li>Xiang, D., 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=Inhibition+of+A/Human/Hubei/3/2005+(H3N2)+influenza+virus+infection+by+silver+nanoparticles+in+vitro+and+in+vivo.">Inhibition of A/Human/Hubei/3/2005 (H3N2) influenza virus infection by silver nanoparticles in vitro and in vivo.</a> International Journal of Nanomedicine. 8 (Issue 1), 4103-4114 (2013).</li><li>Elechiguerra, J. L., 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=Interaction+of+silver+nanoparticles+with+HIV-1.">Interaction of silver nanoparticles with HIV-1.</a> Journal of Nanobiotechnology. 3 (1), 1-10 (2005).</li><li>Nallanthighal, S., 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=Differential+effects+of+silver+nanoparticles+on+DNA+damage+and+DNA+repair+gene+expression+in+Ogg1-deficient+and+wild+type+mice.">Differential effects of silver nanoparticles on DNA damage and DNA repair gene expression in Ogg1-deficient and wild type mice.</a> Nanotoxicology. 11 (8), 1-16 (2017).</li><li>Wen, 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=Acute+toxicity+and+genotoxicity+of+silver+nanoparticle+in+rats.">Acute toxicity and genotoxicity of silver nanoparticle in rats.</a> PLoS One. 12 (9), e0185554 (2017).</li><li>Zhang, R., 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=Silver+nanoparticle+treatment+ameliorates+biliary+atresia+syndrome+in+rhesus+rotavirus+inoculated+mice.">Silver nanoparticle treatment ameliorates biliary atresia syndrome in rhesus rotavirus inoculated mice.</a> Nanomedicine. 13 (3), 1041-1050 (2017).</li><li>Arnold, M., Patton, J. T., McDonald, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culturing,+Storage,+and+Quantification+of+Rotaviruses.">Culturing, Storage, and Quantification of Rotaviruses.</a> Current Protocols in Microbiology. , (2009).</li><li>Liu, X., 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=Silver+nanoparticles+mediate+differential+responses+in+keratinocytes+and+fibroblasts+during+skin+wound+healing.">Silver nanoparticles mediate differential responses in keratinocytes and fibroblasts during skin wound healing.</a> ChemMedChem. 5 (3), 468-475 (2010).</li><li>Zhang, R., 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=Silver+nanoparticles+promote+osteogenesis+of+mesenchymal+stem+cells+and+improve+bone+fracture+healing+in+osteogenesis+mechanism+mouse+model.">Silver nanoparticles promote osteogenesis of mesenchymal stem cells and improve bone fracture healing in osteogenesis mechanism mouse model.</a> Nanomedicine. 11 (8), 1949-1959 (2015).</li><li>Wu, J., Hou, S., Ren, D., Mather, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+properties+of+nanostructured+hydrogel+webs+containing+silver.">Antimicrobial properties of nanostructured hydrogel webs containing silver.</a> Biomacromolecules. 10 (9), 2686-2693 (2009).</li><li>Xu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotoxicity+and+molecular+response+of+silver+nanoparticle+(NP)-based+hydrogel.">Genotoxicity and molecular response of silver nanoparticle (NP)-based hydrogel.</a> Journal of Nanobiotechnology. 10 (1), 16 (2012).</li><li>Dobrzy&#324;ska, M. M., 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=Genotoxicity+of+silver+and+titanium+dioxide+nanoparticles+in+bone+marrow+cells+of+rats+in+vivo.">Genotoxicity of silver and titanium dioxide nanoparticles in bone marrow cells of rats in vivo.</a> Toxicology. 315 (1), 86-91 (2014).</li><li>Mohamed, H. R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+of+TiO+2+nanoparticle-induced+genotoxicity+persistence+and+possible+chronic+gastritis-induction+in+mice.">Estimation of TiO 2 nanoparticle-induced genotoxicity persistence and possible chronic gastritis-induction in mice.</a> Food &amp; Chemical Toxicology. 83 (9), 76-83 (2015).</li></ol>

AgNPs exhibit potent broad-spectrum antibacterial properties and a strong permeability22; additionally, they are used to produce a range of antibacterial medical products23. However, AgNPs can take a long time to clear once they accumulate in organs, and this persistence may lead to toxic effects24,25. A previous study examined the acute toxicity and genotoxicity of AgNPs after a single i.v. injection in a rat exper...

BA is a form of cholestasis characterized by persistent jaundice and has high mortality in the absence of liver transplantation. Viral infections are closely associated with the pathogenesis of BA. The cytomegalovirus, reovirus, and rotavirus have all been suggested as pathogens in BA1,2,3. During the neonatal period, the response of the immature immune system to a viral infection results in immune dysregulation against extra- and intrahepatic bile ducts, leading to biliary epithelial cell apoptosis, inflammatory cell infiltration in the portal area, intrahepatic and extrahepatic bile duct obstruction, and finally, liver fibrosis4,5,6.
The commonly used animal model for BA studies involves the inoculation of a neonatal mouse with the rhesus rotavirus (RRV). The mouse typically develops jaundice after 5 - 6 days, showing a low body weight and acholic stools. The role of the immune response in the disease process is critical, especially for natural killer (NK) cells; the depletion of these cells with anti-NKG2D antibody greatly reduces BA-induced damage7. Furthermore, other cells, including CD4+ T cells, CD8+ T cells, dendritic cells, and regulatory T cells, have all been shown to play roles in the disease8,9,10,11. All data suggest the indispensable nature of the immune system in the course of BA.
Silver nanoparticles (AgNPs) have been demonstrated to have beneficial effects against some infectious diseases, including bacterial infections12 and viral infections13,14,15. However, other than dermatological usage, few studies have used AgNPs in a clinical treatment, mostly because of their potential toxicity. In animal experiments, researchers have generally studied the efficacy of AgNPs administered via oral16 or intravenous methods17. However, no other researchers have studied the efficacy of AgNPs administered via an intraperitoneal (i.p.) injection in neonatal mouse experiments, which is a simple and rapid method leading to a more direct effect on the liver and bile ducts while reducing the toxicity to other systems, such as the immune system. AgNPs have been shown to affect NK cell activity18; therefore, we tested the therapeutic effects of AgNPs administered via i.p. injection in the BA mouse model.

<table>
	<tbody>
		<tr>
			<td>BALB/c mouse</td>
			<td>Guangdong Medical Experimental Animal Center</td>
			<td>SYXK2017-0174</td>
			<td>Animal experiment</td>
		</tr>
		<tr>
			<td>Rhesus rotavirus (RRV)</td>
			<td>ATCC</td>
			<td>ATCC&#160;VR-1739</td>
			<td>Establish biliary atresia mouse model</td>
		</tr>
		<tr>
			<td>MA104 cells</td>
			<td>ATCC</td>
			<td>ATCC&#160;CRL-2378.1</td>
			<td>For laboratory use only</td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Thermo Fisher</td>
			<td>10569010</td>
			<td>Mammalian Cell Culture</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Thermo Fisher</td>
			<td>10099141</td>
			<td>Mammalian Cell Culture</td>
		</tr>
		<tr>
			<td>collagen Type I</td>
			<td>CORNING</td>
			<td>354236</td>
			<td>For research use only</td>
		</tr>
		<tr>
			<td>PBS buffer</td>
			<td>OXOID</td>
			<td>BR0014G</td>
			<td>For washing</td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Sigma</td>
			<td>1310-73-2</td>
			<td>Adjust the PH value</td>
		</tr>
		<tr>
			<td>AgNP</td>
			<td></td>
			<td></td>
			<td>Antibacterial 
			Note: The AgNps was a gift from Prof CM Che. in the Department of Chemistry, the University of Hong Kong.</td>
		</tr>
		<tr>
			<td>Insulin syringe with integrated needle</td>
			<td>BD</td>
			<td>9161635S</td>
			<td>For medical use</td>
		</tr>
		<tr>
			<td>15-mL Centrifuge Tube</td>
			<td>Corning</td>
			<td>430791</td>
			<td>For laboratory use only</td>
		</tr>
		<tr>
			<td>1.5-mL Microcentrifuge Tube</td>
			<td>GEB</td>
			<td>CT0200-B-N</td>
			<td>For laboratory use only</td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Nikon</td>
			<td>ECLIPSE-Ci</td>
			<td>For laboratory use</td>
		</tr>
		<tr>
			<td>Dissecting/Intravital microscope</td>
			<td>Nikon</td>
			<td>SMZ 1000</td>
			<td>For laboratory use</td>
		</tr>
		<tr>
			<td>anti-Mouse NKp46 FITC</td>
			<td>eBioscience</td>
			<td>11-3351</td>
			<td>For research use only</td>
		</tr>
		<tr>
			<td>anti-Mouse CD4 PE-Cyanine5</td>
			<td>eBioscience</td>
			<td>15-0041</td>
			<td>For research use only</td>
		</tr>
		<tr>
			<td>Monoclonal Mouse Anti-Human CD4</td>
			<td>DAKO</td>
			<td>20001673</td>
			<td>For research use only</td>
		</tr>
		<tr>
			<td>anti-NKG2D</td>
			<td>RD</td>
			<td>MAB1547</td>
			<td>For research use only</td>
		</tr>
		<tr>
			<td>BD FACSCanto Flow Cytometer</td>
			<td>BD Biosciences</td>
			<td>FACS Canto Plus</td>
			<td>For laboratory use only</td>
		</tr>
	</tbody>
</table>

a silver nanoparticle method for ameliorating biliary atresia syndrome in mice

Biliary atresia (BA) is a severe type of cholangitis with high mortality in children of which the etiology is still not fully understood. Viral infections may be one possible cause. The typical animal model used for studying BA is established by inoculating a neonatal mouse with a rhesus rotavirus. Silver nanoparticles have been shown to exert antibacterial and antiviral effects; their function in the BA mouse model is evaluated in this study. Currently, in BA animal experiments, the methods used to improve the symptoms of BA mice are generally symptomatic treatments given via food or other drugs. The aim of this study is to demonstrate a new method for ameliorating BA syndrome in mice by the intraperitoneal injection of silver nanoparticles and to provide detailed methods for preparing the silver nanoparticle gel formulation. This method is simple and widely applicable and can be used to research the mechanism of BA, as well as in clinical treatments. Based on the BA mouse model, when the mice exhibit jaundice, the prepared silver nanoparticle gel is injected intraperitoneally to the surface of the lower liver. The survival status is observed, and biochemical indicators and liver histopathology are examined. This method allows a more intuitive understanding of both the establishment of the BA model and novel BA treatments.

Based on the established BA mouse model, the infected neonatal mice were administered an i.p. injection of the prepared AgNP collagen mixture 2x after exhibiting jaundice. Mouse survival was checked for daily, and liver function testing, liver pathology, and flow cytometry were performed. Compared to the untreated control BA mice, the AgNP-treated mice showed reduced jaundice and maintained their normal body weight (Figure 3). The levels of bilirubin metaboli...

Watch this Scientific Journal Video about A Silver Nanoparticle Method for Ameliorating Biliary Atresia Syndrome in Mice at JoVE.com

A Silver Nanoparticle Method for Ameliorating Biliary Atresia Syndrome in Mice

This article describes in detail a method based on silver nanoparticles for ameliorating biliary atresia syndrome in an experimental biliary atresia mouse model. A solid understanding of the reagent preparation process and the neonatal mouse injection technique will help familiarize researchers with the method used in neonatal mouse model studies.

Biliary atresia (BA) is a severe type of cholangitis with high mortality in children of which the etiology is still not fully understood. Viral infections ...

These methods can help answer key questions in biliary atresia. Of BA views about disease, pathogenesis and treatment. The main advantage of this technique is that the neonatal mouse injection technique will help researchers to become familiar with the method for other neonatal mouse model studies.The implications of this technique extend toward the therapy of BA as several nanoparticles of significance Though this method can provide insight into BA treatment, it can also be applied to other systems such as neonatal mouse models of the pathogenesis of this Generally in the way this to this method will because the small size of neonatal mice can result in building of injection process. To establish biliary atresia in a mouse model, load the rhesus rotovirus into a one milliliter insulin syringe equipped with a 29-gauge needle, and intraperitoneally deliver 1.2 times 10 to the 5th plaque-forming units of virus in 20 microliters of Dulbecco's Modified Eagle Medium to each neonate within 24 hours of birth. Then return the neonates to their mother for daily monitoring and weighing.When the neonatal animals begin to exhibit signs of jaundice, load a one milliliter syringe equipped with a 29-gauge needle with 50 microliters of freshly prepared silver nanoparticle collagen solution per animal and use the ring finger to press the leg of one jaundiced animal obliquely over the right thigh. Slowly introduce the needle at a 15 degree angle into the peritoneum, and upon reaching the surface of the lower edge of the liver, inject the silver nanoparticle collagen mixture. When all of the mixture has been injected, slowly remove the needle and allow the animal to rest for ten minutes to allow the collagen to solidify and to prevent the mother from licking the injection site.Then return the mice to their cages for daily monitoring. On day nine or twelve after inoculation, sterilize the upper and lower abdomen with 75%ethanol, and immobilize the limbs of an infected mouse. Open the skin, muscle and peritoneum along the midline to the xyphoid, and use a sterile cotton swab to remove the gastrointestinal tract to fully expose the diaphragm.Insert the needle of an unloaded one milliliter insulin syringe into the left ventricle of the heart, and slowly retract the syringe plunger to obtain the maximum blood volume. Transfer the blood to a 1.5 milliliter tube for a 30 minute incubation at room temperature and pellet the red blood cells by centrifugation. Then collect the serum for later analysis.For extrahepatic cholangiography, use a cotton swab to fully expose the liver, gallbladder and extrahepatic bile ducts. After photographing the appearance of the liver and bile ducts under a dissecting microscope, use ophthalmic forceps to gently grasp the bottom of the gallbladder and slowly insert the needle of a one milliliter syringe loaded with methylene blue solution into the gallbladder cavity. Grasping the needle with the forceps, slowly infuse 10-20 microliters of methylene blue.When the dye passes through the extrahepatic bile ducts to the jejunum, obtain another photograph of the tissue. Then for hematoxylin and eosin staining, harvest the liver from each animal for overnight fixation in 10%formalin. The next morning, embed the samples in paraffin for sectioning, followed by de-waxing re-hydration with an ethanol series and hematoxylin and eosin staining according to standard histopathological analysis protocols.Compared to untreated biliary atresia mice, silver nanoparticle treated animals demonstrate reduced jaundice and maintain their normal body weight. The levels of bilirubin metabolism and hepatic transaminase drop to normal control values, suggesting that the silver nanoparticles greatly improve liver function. Extrahepatic cholangiography with methylene blue staining confirms bile duct patency after silver nanoparticle treatment, and H&E staining reveals a significant decrease in inflammatory cell infiltration in the hepatic portal area of mice treated with silver nanoparticles compared to controls.Flow site and metric analysis indicates the presence of significantly fewer NK cells in the livers of silver nanoparticle treated animals compared to untreated rhesus rotavirus infected mice. Further immunohistochemical staining reveals a substantially reduced expression of NK cell marker staining in the portal triad of silver nanoparticle treated mice compared to rhesus rotavirus infected animals. While attempting this procedure, it's important to remember that practice will give best results.Following this procedure, as methods like conjugating silver nanoparticles to other things can be performed to answer additional questions about the activity of lesions in the treatments of BA in mouse models. is development. This technique provided a way for researchers in the field of gastroenterology to BA or other diseases such as cholestasis in the liver.Don't forget that working with research rotavirus can be extremely hazardous, and thus precautions such as wearing the appropriate personal protective equipment should always be taken when performing this procedure.

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