Name: direct intrabronchial administration to improve the selective agent deposition within the mouse lung
Uploaded: 2019-05-20T14:00:00
Description: Watch this Scientific Journal Video about Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung at JoVE.com

This work was funded by NHLBI grant HL125371 to E.P.S. and by DOD (CDMRP) grant W81XWH-17-1-0051 to Y.Y.

All animal protocols have been approved by the University of Colorado Denver Institutional Animal Care and Use Committee (IACUC). All procedures described below (sections 4&#8211;7) have been optimized using both male and female C57BL/6 mice. This approach has been validated using mice ranging from 19&#8211;40 g in body weight.
1. Creation of platform for IB administration
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	<li>Bend the bookend from the original 90&#176; angle between the basal wing and standing wing to 70&#176; (<stron...

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	<li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> Journal of Applied Physiology. 106 (3), 984-987 (2009).</li><li>Thomas, 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=Endotracheal+intubation+in+mice+via+direct+laryngoscopy+using+an+otoscope.">Endotracheal intubation in mice via direct laryngoscopy using an otoscope.</a> Journal of Visualized Experiments. (86), (2014).</li><li>Vandivort, T. C., An, D., Parks, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Improved+Method+for+Rapid+Intubation+of+the+Trachea+in+Mice.">An Improved Method for Rapid Intubation of the Trachea in Mice.</a> Journal of Visualized Experiments. (108), 53771 (2016).</li><li>McGovern, T. K., Robichaud, A., Fereydoonzad, L., Schuessler, T. F., Martin, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+respiratory+system+mechanics+in+mice+using+the+forced+oscillation+technique.">Evaluation of respiratory system mechanics in mice using the forced oscillation technique.</a> Journal of Visualized Experiments. (75), e50172 (2013).</li><li>Halbower, A. C., Mason, R. J., Abman, S. H., Tuder, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Agarose+infiltration+improves+morphology+of+cryostat+sections+of+lung.">Agarose infiltration improves morphology of cryostat sections of lung.</a> Laboratory Investigation. 71 (1), 149-153 (1994).</li><li>Thrall, R. S., McCormick, J. R., Jack, R. M., McReynolds, R. A., Ward, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bleomycin-induced+pulmonary+fibrosis+in+the+rat:+inhibition+by+indomethacin.">Bleomycin-induced pulmonary fibrosis in the rat: inhibition by indomethacin.</a> American Journal of Pathology. 95 (1), 117-130 (1979).</li><li>Matute-Bello, G., Frevert, C. W., Martin, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+acute+lung+injury.">Animal models of acute lung injury.</a> American Journal of Physiology Lung Cellular and Molecular Physiology. 295 (3), L379-L399 (2008).</li><li>Del Sorbo, 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=Intratracheal+Administration+of+Small+Interfering+RNA+Targeting+Fas+Reduces+Lung+Ischemia-Reperfusion+Injury.">Intratracheal Administration of Small Interfering RNA Targeting Fas Reduces Lung Ischemia-Reperfusion Injury.</a> Criticial Care Medicine. 44 (8), e604-e613 (2016).</li><li>McLemore, T. 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=Novel+intrapulmonary+model+for+orthotopic+propagation+of+human+lung+cancers+in+athymic+nude+mice.">Novel intrapulmonary model for orthotopic propagation of human lung cancers in athymic nude mice.</a> Cancer Research. 47 (19), 5132-5140 (1987).</li><li>Vertrees, R. 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=Development+of+a+human+to+murine+orthotopic+xenotransplanted+lung+cancer+model.">Development of a human to murine orthotopic xenotransplanted lung cancer model.</a> Journal of Investigative Surgery. 13 (6), 349-358 (2000).</li></ol>

Lung injury has been classically modeled in rodents using IT administration of injurious agents such as BLM6. Such IT administration, however, only leads to patchy injury, reflecting the nontargeted nature of lung delivery with this approach7. These limitations of modeling lung injury are instructive challenges faced when attempting the IT delivery of non-injurious experimental agents, such as drugs, siRNA, or cellular therapies.
In this report, ...

Direct pulmonary administration of experimental agents in mice allows for the study of lung immune responses, acute lung injury, and lung fibrosis. Direct pulmonary administration is typically performed via intratracheal (IT) instillation, as described previously1,2,3. However, this approach is nonselective, affecting both lungs in a nontargeted and often asymmetric fashion.&#160; Experimental modeling of lung injury may benefit from the ability to selectively target one specific lung, allowing for use of the contralateral lung as a control. Conversely, accurate modeling of human diffuse lung diseases benefits from symmetric distribution of experimental agents to the bilateral lung parenchyma.
The overall goal of this report is to describe a method for selective delivery of experimental agents to the left or right lung of a mouse (Figure 1). This intrabronchial (IB) administration approach allows for unilateral treatment of a mouse lung and can be easily adapted to ensure equal delivery of an agent to the bilateral mainstem bronchi. By using IB administration to deliver larger doses of experimental agents to the larger right lung and smaller volumes to the smaller left lung (i.e., dose-adjusted IB administration), demonstrated in this report is an improvement in the homogeneity of pulmonary delivery of experimental agents, optimizing the model of diffuse lung injury in mice. As such, this report may hold value for investigators seeking to either unilaterally administer experimental agents to mice or improve the symmetry of drug deposition in both lungs.

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			<td height="21" style="height:21px;width:280px;">22 G shielded IV Catheter</td>
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			<td height="21" style="height:21px;width:280px;">Bleomycin</td>
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			<td height="42" style="height:42px;width:280px;">Compact Mini rodent anesthesia machine&#160;</td>
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			<td height="21" style="height:21px;width:280px;">FITC-dextran</td>
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			<td height="42" style="height:42px;width:280px;">Isoflurane</td>
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			<td height="42" style="height:42px;width:280px;">LED-30W Fiber Optic Dual Gooseneck Lights Microscope Illuminator</td>
			<td style="width:139px;">AmScope</td>
			<td style="width:140px;">LED-30W</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Low temperature cautery with fine tip&#160;</td>
			<td style="width:139px;">Bovie</td>
			<td style="width:140px;">AA02</td>
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			<td height="21" style="height:21px;width:280px;">Precisionglide needle, 18G x 1&#34;</td>
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			<td>Beveled tip, 12 mm in length&#160;</td>
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			<td height="21" style="height:21px;width:280px;">Xylazine</td>
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			<td style="width:140px;">NDC 59399-110-20</td>
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			<td height="21" style="height:21px;width:280px;">Zatamine</td>
			<td>VetOne</td>
			<td style="width:140px;">NDC 13985-702-10&#160;</td>
			<td>Ketamine</td>
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direct intrabronchial administration to improve the selective agent deposition within the mouse lung

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced pulmonary fibrosis.&#160; However, distribution of intratracheally-administered agents to the distal mouse lung is often asymmetric, with lung parenchymal concentrations increased in the smaller (but equally accessible) left lung of the mouse.&#160; Described in this report is a novel intrabronchial (IB) approach to cannulate the left and/or right lungs of living mice non-operatively.&#160; It is also demonstrated how this approach can be used to selectively administer agents to one lung or adapted (via dose-adjusted IB delivery) to improve the left-right symmetry of lung delivery of experimental agents, thereby improving models of diffuse lung disease such as bleomycin-induced pulmonary fibrosis.

Selective IB intubation targets specific lobes (right lung) or basilar segments (left lung).
IB administration of EBD to the right lung was performed as described in section 5.1. After completion of the experiment, mice were administered a lethal dose of intraperitoneal ketamine/xylazine, and lungs were harvested for demonstration of EBD distribution (Figure 4A, right). Gross appear...

Watch this Scientific Journal Video about Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung at JoVE.com

Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung

Intratracheal (IT) administration of experimental agents in mice often results in asymmetric delivery to the distal lungs.&#160; In this report, we describe a direct intrabronchial (IB) approach to cannulate each lung in living mice non-operatively.&#160; This approach can be used to selectively administer agents to one lung or may be adapted to improve the symmetric agent delivery to both lungs.

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced ...

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