Gli autori ringraziano il professor Yu Feng, la dott.ssa Jenna Briddell, Ian Woodward e Lucas Attia per le loro utili discussioni.

<ol>
	<li>Goel, A., Baboota, S., Sahni, J. K., Ali, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploring+targeted+pulmonary+delivery+for+treatment+of+lung+cancer.">Exploring targeted pulmonary delivery for treatment of lung cancer.</a> International Journal of Pharmaceutical Investigation. 3 (1), 8-14 (2013).</li><li>Kleinstreuer, C., Zhang, Z., Li, Z., Roberts, W. L., Rojas, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+methodology+for+targeting+drug-aerosols+in+the+human+respiratory+system.">A new methodology for targeting drug-aerosols in the human respiratory system.</a> International Journal of Heat and Mass Transfer. 51 (23), 5578-5589 (2008).</li><li>Feng, Y., Chen, X., Yang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+In+Silico+Investigation+of+a+Lobe-Specific+Targeted+Pulmonary+Drug+Delivery+Method.">An In Silico Investigation of a Lobe-Specific Targeted Pulmonary Drug Delivery Method.</a> Design of Medical Devices Conference. , (2018).</li><li>Marple, V. 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=Next+generation+pharmaceutical+impactor+(a+new+impactor+for+pharmaceutical+inhaler+testing).+Part+I:+Design.">Next generation pharmaceutical impactor (a new impactor for pharmaceutical inhaler testing). Part I: Design.</a> Journal of Aerosol Medicine. 16 (3), 283-299 (2003).</li><li>Feng, Y., Zhao, J., Chen, X., Lin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+In+Silico+Subject-Variability+Study+of+Upper+Airway+Morphological+Influence+on+the+Airflow+Regime+in+a+Tracheobronchial+Tree.">An In Silico Subject-Variability Study of Upper Airway Morphological Influence on the Airflow Regime in a Tracheobronchial Tree.</a> Bioengineering. 4 (4), 90 (2017).</li><li>Huynh, B. 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=The+Development+and+Validation+of+an+In+Vitro+Airway+Model+to+Assess+Realistic+Airway+Deposition+and+Drug+Permeation+Behavior+of+Orally+Inhaled+Products+Across+Synthetic+Membranes.">The Development and Validation of an In Vitro Airway Model to Assess Realistic Airway Deposition and Drug Permeation Behavior of Orally Inhaled Products Across Synthetic Membranes.</a> Journal of Aerosol Medicine and Pulmonary Drug Delivery. 31 (2), 103-108 (2018).</li><li>Lizal, F., Elcner, J., Hopke, P. K., Jedelsky, J., Jicha, M. <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+realistic+human+airway+model.">Development of a realistic human airway model.</a> Proceedings of the Institution of Mechanical Engineers, Part H: Journal of Engineering in Medicine. 226 (3), 197-207 (2011).</li><li>Wei, X., Hindle, M., Delvadia, R. R., Byron, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+Vitro+Tests+for+Aerosol+Deposition.+V:+Using+Realistic+Testing+to+Estimate+Variations+in+Aerosol+Properties+at+the+Trachea.">In Vitro Tests for Aerosol Deposition. V: Using Realistic Testing to Estimate Variations in Aerosol Properties at the Trachea.</a> Journal of Aerosol Medicine and Pulmonary Drug Delivery. 30 (5), 339-348 (2017).</li><li>Kolewe, E. L., Feng, Y., Fromen, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Realizing+Lobe-Specific+Aerosol+Targeting+in+a+3D-Printed+In+Vitro+Lung+Model.">Realizing Lobe-Specific Aerosol Targeting in a 3D-Printed In Vitro Lung Model.</a> Journal of Aerosol Medicine and Pulmonary Drug Delivery. , (2020).</li><li>Sul, B., 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=Assessing+Airflow+Sensitivity+to+Healthy+and+Diseased+Lung+Conditions+in+a+Computational+Fluid+Dynamics+Model+Validated+In+Vitro.">Assessing Airflow Sensitivity to Healthy and Diseased Lung Conditions in a Computational Fluid Dynamics Model Validated In Vitro.</a> Journal of Biomechanical Engineering. 140 (5), (2018).</li><li>Martonen, T. B., Katz, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deposition+Patterns+of+Polydisperse+Aerosols+Within+Human+Lungs.">Deposition Patterns of Polydisperse Aerosols Within Human Lungs.</a> Journal of Aerosol Medicine. 6 (4), 251-274 (1993).</li><li>Nahar, 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=In+vitro,+in+vivo+and+ex+vivo+models+for+studying+particle+deposition+and+drug+absorption+of+inhaled+pharmaceuticals.">In vitro, in vivo and ex vivo models for studying particle deposition and drug absorption of inhaled pharmaceuticals.</a> European Journal of Pharmaceutical Sciences. 49 (5), 805-818 (2013).</li><li>Nichols, S. C., 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=A+Multi-laboratory+in+Vitro+Study+to+Compare+Data+from+Abbreviated+and+Pharmacopeial+Impactor+Measurements+for+Orally+Inhaled+Products:+a+Report+of+the+European+Aerosol+Group+(EPAG).">A Multi-laboratory in Vitro Study to Compare Data from Abbreviated and Pharmacopeial Impactor Measurements for Orally Inhaled Products: a Report of the European Aerosol Group (EPAG).</a> AAPS PharmSciTech. 17 (6), 1383-1392 (2016).</li><li>Yoshida, H., Kuwana, A., Shibata, H., Izutsu, K. I., Goda, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+Aerodynamic+Particle+Size+Distribution+Between+a+Next+Generation+Impactor+and+a+Cascade+Impactor+at+a+Range+of+Flow+Rates.">Comparison of Aerodynamic Particle Size Distribution Between a Next Generation Impactor and a Cascade Impactor at a Range of Flow Rates.</a> AAPS PharmSciTech. 18 (3), 646-653 (2017).</li><li>Feng, Y., 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=An+in+silico+inter-subject+variability+study+of+extra-thoracic+morphology+effects+on+inhaled+particle+transport+and+deposition.">An in silico inter-subject variability study of extra-thoracic morphology effects on inhaled particle transport and deposition.</a> Journal of Aerosol Science. 123, 185-207 (2018).</li><li>Kleinstreuer, C., Seelecke, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhaler+system+for+targeted+maximum+drug-aerosol+delivery.">Inhaler system for targeted maximum drug-aerosol delivery.</a> United States patent. , (2005).</li><li>. How Medical 3D Printing is Gaining Ground in Top Hospitals Available from: <a target="_blank" href="https://www.materialise.com/en/blog/3d-printing-hospitals">https://www.materialise.com/en/blog/3d-printing-hospitals</a> (2019)</li><li>Weber, P. W., Price, O. T., McClellan, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demographic+Variability+of+Inhalation+Mechanics:+A+Review.">Demographic Variability of Inhalation Mechanics: A Review.</a> Defense Threat Reduction Agency. , (2016).</li><li>Jiang, Y. Y., Xu, X., Su, H. L., Liu, D. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender-related+difference+in+the+upper+airway+dimensions+and+hyoid+bone+position+in+Chinese+Han+children+and+adolescents+aged+6-18+years+using+cone+beam+computed+tomography.">Gender-related difference in the upper airway dimensions and hyoid bone position in Chinese Han children and adolescents aged 6-18 years using cone beam computed tomography.</a> Acta Odontologica Scandinavica. 73 (5), 391-400 (2015).</li><li>Martin, S. E., Mathur, R., Marshall, I., Douglas, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+age,+sex,+obesity+and+posture+on+upper+airway+size.">The effect of age, sex, obesity and posture on upper airway size.</a> European Respiratory Journal. 10 (9), 2087 (1997).</li><li>Xi, J., Longest, P. W., Martonen, T. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+the+laryngeal+jet+on+nano-+and+microparticle+transport+and+deposition+in+an+approximate+model+of+the+upper+tracheobronchial+airways.">Effects of the laryngeal jet on nano- and microparticle transport and deposition in an approximate model of the upper tracheobronchial airways.</a> Journal of Applied Physiology. 104 (6), 1761-1777 (2008).</li><li>Zhao, J., Feng, Y., Fromen, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glottis+motion+effects+on+the+particle+transport+and+deposition+in+a+subject-specific+mouth-to-trachea+model:+A+CFPD+study.">Glottis motion effects on the particle transport and deposition in a subject-specific mouth-to-trachea model: A CFPD study.</a> Computers in Biology and Medicine. 116, 103532 (2020).</li><li>Kim, S. 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=Chronic+obstructive+pulmonary+disease:+lobe-based+visual+assessment+of+volumetric+CT+by+Using+standard+images--comparison+with+quantitative+CT+and+pulmonary+function+test+in+the+COPDGene+study.">Chronic obstructive pulmonary disease: lobe-based visual assessment of volumetric CT by Using standard images--comparison with quantitative CT and pulmonary function test in the COPDGene study.</a> Radiology. 266 (2), 626-635 (2013).</li><li>. The Cancer Imaging Archive Available from: <a target="_blank" href="https://www.cancerimagingarchive.net/">https://www.cancerimagingarchive.net/</a> (2020)</li><li>Li, A., Ahmadi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computer+Simulation+of+Deposition+of+Aerosols+in+a+Turbulent+Channel+Flow+with+Rough+Walls.">Computer Simulation of Deposition of Aerosols in a Turbulent Channel Flow with Rough Walls.</a> Aerosol Science and Technology. 18 (1), 11-24 (1993).</li><li>Khalili, S. F., Ghanbarzadeh, S., Nokhodchi, A., Hamishehkar, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+different+coating+materials+on+the+prevention+of+powder+bounce+in+the+next+generation+impactor.">The effect of different coating materials on the prevention of powder bounce in the next generation impactor.</a> Research in Pharmaceutical Sciences. 13 (3), 283-287 (2018).</li><li>Galliger, Z., Vogt, C. D., Panoskaltsis-Mortari, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+bioprinting+for+lungs+and+hollow+organs.">3D bioprinting for lungs and hollow organs.</a> Translational Research. 211, 19-34 (2019).</li><li>Schwarz, K., Biller, H., Windt, H., Koch, W., Hohlfeld, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+exhaled+particles+from+the+healthy+human+lung--a+systematic+analysis+in+relation+to+pulmonary+function+variables.">Characterization of exhaled particles from the healthy human lung--a systematic analysis in relation to pulmonary function variables.</a> Journal of Aerosol Medicine and Pulmonary Drug Delivery. 23 (6), 371-379 (2010).</li><li>Patton, J. S., Byron, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhaling+medicines:+delivering+drugs+to+the+body+through+the+lungs.">Inhaling medicines: delivering drugs to the body through the lungs.</a> Nature Reviews Drug Discovery. 6 (1), 67-74 (2007).</li><li>Zhang, Z., Kleinstreuer, C., Kim, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+micron-size+particle+inhalation+and+deposition+in+a+triple+bifurcation+lung+airway+model.">Cyclic micron-size particle inhalation and deposition in a triple bifurcation lung airway model.</a> Journal of Aerosol Science. 33 (2), 257-281 (2002).</li><li>Ju, Y., 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=Engineering+of+Nebulized+Metal-Phenolic+Capsules+for+Controlled+Pulmonary+Deposition.">Engineering of Nebulized Metal-Phenolic Capsules for Controlled Pulmonary Deposition.</a> Advanced Science. 7 (6), 1902650 (2020).</li></ol>

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L'attuale dispositivo all'avanguardia per il test farmaceutico polmonare di una dose completa di inalazione è il Next Generator Impactor (NGI), che misura il diametro aerodinamico di un aerosol4. Questi dati di dimensionamento vengono quindi utilizzati per prevedere la generazione polmonare in cui l'aerosol si depositerà sulla base di una correlazione sviluppata per un maschio adultosano 11. Sfortunatamente, questo metodo è limitato nella sua capacità di valutare le dif...

Molte malattie polmonari come il cancro ai polmoni e la broncopneumopatia cronica ostruttiva (BPCO) presentano differenze regionali nelle caratteristiche della malattia; tuttavia, mancano tecniche terapeutiche disponibili per indirizzare la somministrazione di farmaci solo alle regioni masmato del polmone1. Molteplici modelli di fluidodinamica computazionale (CFD) hanno dimostrato che è possibile modulare i profili di deposizione dei farmaci identificando specifiche snellenel polmone 2,3. Nello stesso laboratorio sono in corso lo sviluppo di inalatori e adattatori per tubi endotracheali (ET) con capacità di targeting regionali per controllare la distribuzione di aerosol nelle regioni polmonari mante. L'estensione di questi principi all'uso clinico è limitata dall'attuale capacità di test preclinici. La posizione precisa in cui un farmaco si deposita all'interno del polmone è nota per essere il miglior predittore di efficacia; tuttavia, le attuali valutazioni farmaceutiche delle terapie inalabili sono più spesso previste utilizzando correlazioni in vitro-in vivo della dimensione delle particelle per approssimare semplicemente la deposizione4. Questa tecnica non consente alcuna analisi spaziale per determinare gli effetti delle diverse geometrie delle vie aeree sulla distribuzione regionale attraverso i vari lobi del polmone. Inoltre, questo test manca di geometrie polmonari anatomicamente accurate, che i ricercatori hanno dimostrato possono avere un impatto significativo sui profili di deposizione5. Sono stati compiuti alcuni sforzi per incorporare geometrie polmonari specifiche del paziente nei protocolli di prova attraverso l'aggiunta delle vie aeree superiori; tuttavia, la maggior parte di questi approcci campiona l'erogazione di aerosol a varie generazioni del polmone piuttosto che a ciascun lobopolmonare 6,7,8. Il seguente protocollo presenta un metodo ad alta produttività per generare modelli polmonari specifici del paziente con la capacità di quantificare la deposizione relativa di particelle in ciascuno dei cinque lobi delpolmone 9.
I polmoni modello anatomicamente accurati sono generati dalla stampa 3D di scansioni di tomografia computerizzata (CT) del paziente. Se utilizzate in combinazione con un sistema di flusso facilmente assemblabile, le portate relative attraverso ciascuno dei lobi del polmone modello possono essere controllate in modo indipendente e su misura per imitare quelle di diversi stati demografici e / o di malattia del paziente. Con questo metodo, i ricercatori possono testare l'efficacia di potenziali metodi terapeutici in una geometria polmonare rilevante e correlare le prestazioni di ogni metodo con la progressione della morfologia masabile. Qui, due progetti di dispositivi sviluppati nel nostro laboratorio vengono testati per la loro capacità di aumentare la deposizione in un lobo polmonare desiderato controllando la posizione del rilascio di aerosol in bocca o trachea. Questo protocollo ha anche il potenziale per influire in modo significativo sullo sviluppo di procedure personalizzate per i pazienti facilitando la rapida previsione dell'efficacia del trattamento in un polmone modello specifico per i dati di TAC di quel paziente.

<table>
	<tbody>
		<tr>
			<td>1/4&#34; Plastic Barbed Tube Fitting</td>
			<td>McMaster Carr</td>
			<td>5372K111</td>
			<td></td>
		</tr>
		<tr>
			<td>10 um Filter Paper</td>
			<td>Fisher</td>
			<td>1093-110</td>
			<td></td>
		</tr>
		<tr>
			<td>1um Fluorescent Polystyrene Particles</td>
			<td>Polysciences</td>
			<td>15702-10</td>
			<td></td>
		</tr>
		<tr>
			<td>1um Non-Fluorescent Polystyrene Particles</td>
			<td>Polysciences</td>
			<td>8226</td>
			<td></td>
		</tr>
		<tr>
			<td>2-Propanol</td>
			<td>Fisher</td>
			<td>A516-4</td>
			<td>Referred to in protocol as &#34;IPA&#34;</td>
		</tr>
		<tr>
			<td>3/8&#34; Plastic Barbed Tube Fitting</td>
			<td>McMaster Carr</td>
			<td>5372K117</td>
			<td></td>
		</tr>
		<tr>
			<td>Air Flow Meter (1 - 280 mL/min)</td>
			<td>McMaster Carr</td>
			<td>41695K32</td>
			<td>Referred to in protocol as &#34;flow meter&#34;</td>
		</tr>
		<tr>
			<td>Carbon M1 3D Printer</td>
			<td>Carbon 3D</td>
			<td></td>
			<td>https://www.carbon3d.com/, Associated software referred to in protocol as &#34;slicing software&#34;</td>
		</tr>
		<tr>
			<td>Collison Jet Nebulizer</td>
			<td>CH Technologies</td>
			<td>ARGCNB0008 (CN-25)</td>
			<td>6 Jet MRE style horizontal collision with glass jar, Referred to in protocol as &#34;nebulizer&#34;, http://chtechusa.com/Manuals/MRE_Collison_Manual.pdf</td>
		</tr>
		<tr>
			<td>Convection Oven</td>
			<td>Yamato</td>
			<td>DKN602</td>
			<td></td>
		</tr>
		<tr>
			<td>Copley Critical Flow Controller TPK2000 Reve 120V</td>
			<td>MSP Corp</td>
			<td>0001-01-9810</td>
			<td>Referred to in protocol as &#34;flow controller&#34;</td>
		</tr>
		<tr>
			<td>Copley High Capacity Pump Model HCP5</td>
			<td>MSP Corp</td>
			<td>0001-01-9982</td>
			<td>Referred to in protocol as &#34;vacuum pump&#34;</td>
		</tr>
		<tr>
			<td>Cytation</td>
			<td>BioTek</td>
			<td>CYT5MPV</td>
			<td>Multifunctional Spectrophotometer/Fluorescent imager equiped with 4x/20x/40x objectives and DAPI/GFP/TexasRed laser/filter cubes</td>
		</tr>
		<tr>
			<td>EPU40 Resin</td>
			<td>Carbon 3D</td>
			<td></td>
			<td>https://www.carbon3d.com/materials/epu-elastomeric-polyurethane/, Referred to in protocol as &#34;soft resin&#34;</td>
		</tr>
		<tr>
			<td>Filter for vacuum pump</td>
			<td>Whatman</td>
			<td>6722-5000</td>
			<td></td>
		</tr>
		<tr>
			<td>Flow Meter Model DFM 2000</td>
			<td>MSP Corp</td>
			<td>0001-01-8764</td>
			<td>Referred to in protocol as &#34;electronic flow meter&#34;</td>
		</tr>
		<tr>
			<td>ImageJ Software</td>
			<td>ImageJ</td>
			<td></td>
			<td>https://imagej.nih.gov/ij/download.html</td>
		</tr>
		<tr>
			<td>Inline Air Flow Control Valve (Push-to-Connect)</td>
			<td>McMaster Carr</td>
			<td>62005K333</td>
			<td>Referred to in protocol as &#34;valve&#34;</td>
		</tr>
		<tr>
			<td>Inline Filter Devices</td>
			<td>Whatman</td>
			<td>WHA67225000</td>
			<td></td>
		</tr>
		<tr>
			<td>Marine-Grade Plywood Sheet</td>
			<td>McMaster Carr</td>
			<td>62005K333</td>
			<td>Referred to in protocol as &#34;wooden board&#34;</td>
		</tr>
		<tr>
			<td>Materialise Mimics Software</td>
			<td>Materialise</td>
			<td></td>
			<td>https://www.materialise.com/en/medical/mimics-innovation-suite, Referred to in protocol as &#34;CT scan software&#34;</td>
		</tr>
		<tr>
			<td>Meshmixer Software</td>
			<td>Autodesk</td>
			<td></td>
			<td>http://www.meshmixer.com/, Referred to in protocol as &#34;mesh editing software&#34;</td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Fisher</td>
			<td>A454-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Opticure LED Cube</td>
			<td>APM Technica</td>
			<td>102843</td>
			<td>Referred to in protocol as &#34;UV oven&#34;</td>
		</tr>
		<tr>
			<td>PR25 Resin</td>
			<td>Carbon 3D</td>
			<td></td>
			<td>https://www.carbon3d.com/materials/uma-urethanemethacrylate, /Referred to in protocol as &#34;hard resin&#34;</td>
		</tr>
		<tr>
			<td>PVC Tube for Chemicals</td>
			<td>McMaster Carr</td>
			<td>5231K161</td>
			<td>1/4&#34; ID</td>
		</tr>
		<tr>
			<td>Screws</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SolidWorks Software</td>
			<td>Dassault Syst&#232;mes&#160;SolidWorks Corporation</td>
			<td></td>
			<td>https://www.solidworks.com/, Referred to in protocol as &#34;3D modeling software&#34;</td>
		</tr>
		<tr>
			<td>Straight Flow Rectangular Manifold</td>
			<td>McMaster Carr</td>
			<td>1125T31</td>
			<td></td>
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evaluating regional pulmonary deposition using patient-specific 3d printed lung models

Lo sviluppo di terapie mirate per le malattie polmonari è limitato dalla disponibilità di metodi di test preclinici con la capacità di prevedere l'erogazione regionale di aerosol. Sfruttando la stampa 3D per generare modelli polmonari specifici del paziente, descriviamo la progettazione di una configurazione sperimentale in vitro ad alta produttività per quantificare la deposizione polmonare lobulare. Questo sistema è realizzato con una combinazione di componenti stampati in commercio e stampati in 3D e consente di controllare in modo indipendente la portata attraverso ogni lobo del polmone. La consegna di aerosol fluorescenti a ciascun lobo viene misurata utilizzando la microscopia a fluorescenza. Questo protocollo ha il potenziale per promuovere la crescita della medicina personalizzata per le malattie respiratorie attraverso la sua capacità di modellare una vasta gamma di dati demografici dei pazienti e stati di malattia. Sia la geometria del modello polmonare stampato in 3D che l'impostazione del profilo del flusso d'aria possono essere facilmente modulate per riflettere i dati clinici per pazienti con età, razza e sesso variabili. I dispositivi di somministrazione dei farmaci clinicamente rilevanti, come il tubo endotracheale mostrato qui, possono essere incorporati nella configurazione del test per prevedere in modo più accurato la capacità di un dispositivo di indirizzare l'erogazione terapeutica in una regione masabile del polmone. La versatilità di questa configurazione sperimentale permette di personalizzarla per riflettere una moltitudine di condizioni di inalazione, esaltando il rigore dei test terapeutici preclinici.

Particelle in questo intervallo di dimensioni (1-5 μm) e condizioni di flusso (1-10 L/min) seguono le linee del flusso fluido in base sia al loro numero teorico di Stokes che ai dati in vivo; pertanto, in assenza di un dispositivo di consegna mirato, le particelle rilasciate nel modello polmonare dovrebbero depositarsi in base alla percentuale del flusso d'aria totale dirottato su ciascun lobo. Le quantità relative di consegna di particelle a ciascun lobo possono quindi essere confrontate con i dati clinici sulla porta...

Watch this Scientific Journal Video about Evaluating Regional Pulmonary Deposition using Patient-Specific 3D Printed Lung Models at JoVE.com

Evaluating Regional Pulmonary Deposition using Patient-Specific 3D Printed Lung Models

Presentiamo un metodo in vitro ad alta produttività per quantificare la deposizione polmonare regionale a livello di lobo utilizzando modelli polmonari stampati in 3D derivati dalla TAC con profili di flusso d'aria tonnibili.

Valutazione della deposizione polmonare regionale utilizzando modelli polmonari stampati in 3D specifici del paziente

Development of targeted therapies for pulmonary diseases is limited by the availability of preclinical testing methods with the ability to predict ...

evaluating-regional-pulmonary-deposition-using-patient-specific-3d

Universidad de Cartagena UDEC

Research

JoVE Journal

Bioengineering

4.2K Views.  University of Delaware. Presentiamo un metodo in vitro ad alta produttività per quantificare la deposizione polmonare regionale a livello di lobo utilizzando modelli polmonari stampati in 3D derivati dalla TAC con profili di flusso d'aria tonnibili.

Video: Evaluating Regional Pulmonary Deposition using Patient-Specific 3D Printed Lung Models

Patient-specific Modeling of the Heart: Estimation of Ventricular Fiber Orientations

Patient-specific simulations of heart (dys)function aimed at personalizing cardiac therapy are hampered by the absence of in vivo imaging technology for clinically acquiring myocardial fiber orientations. The objective of this project was to develop a methodology to estimate cardiac fiber orientations from in vivo images of patient heart geometries. An accurate representation of ventricular geometry and fiber orientations was reconstructed, respectively, from high-resolution ex vivo structural magnetic resonance (MR) and diffusion tensor (DT) MR images of a normal human heart, referred to as the atlas. Ventricular geometry of a patient heart was extracted, via semiautomatic segmentation, from an in vivo computed tomography (CT) image. Using image transformation algorithms, the atlas ventricular geometry was deformed to match that of the patient. Finally, the deformation field was applied to the atlas fiber orientations to obtain an estimate of patient fiber orientations. The accuracy of the fiber estimates was assessed using six normal and three failing canine hearts. The mean absolute difference between inclination angles of acquired and estimated fiber orientations was 15.4 &deg;. Computational simulations of ventricular activation maps and pseudo-ECGs in sinus rhythm and ventricular tachycardia indicated that there are no significant differences between estimated and acquired fiber orientations at a clinically observable level.The new insights obtained from the project will pave the way for the development of patient-specific models of the heart that can aid physicians in personalized diagnosis and decisions regarding electrophysiological interventions.

A methodology to estimate ventricular fiber orientations from in vivo images of patient heart geometries for personalized modeling is described. Validation of the methodology performed using normal and failing canine hearts demonstrate that that there are no significant differences between estimated and acquired fiber orientations at a clinically observable level.

Paziente-specifica modellazione del Cuore: Stima di orientamento delle fibre ventricolari

Patient-specific  simulations of heart (dys)function aimed at personalizing cardiac therapy are hampered  by the absence of in vivo imaging technology for ...

Ricerca

Bioingegneria

Longitudinal Measurement of Extracellular Matrix Rigidity in 3D Tumor Models Using Particle-tracking Microrheology

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and modified as part of a set of complex, two-way mechanosensitive interactions. While the development of biologically-relevant 3D tumor models have facilitated mechanistic studies on the impact of matrix rheology on tumor growth, the inverse problem of mapping changes in the mechanical environment induced by tumors remains challenging. Here, we describe the implementation of particle-tracking microrheology (PTM) in conjunction with 3D models of pancreatic cancer as part of a robust and viable approach for longitudinally monitoring physical changes in the tumor microenvironment, in situ. The methodology described here integrates a system of preparing in vitro 3D models embedded in a model extracellular matrix (ECM) scaffold of Type I collagen with fluorescently labeled probes uniformly distributed for position- and time-dependent microrheology measurements throughout the specimen. In vitro tumors are plated and probed in parallel conditions using multiwell imaging plates. Drawing on established methods, videos of tracer probe movements are transformed via the Generalized Stokes Einstein Relation (GSER) to report the complex frequency-dependent viscoelastic shear modulus, G*(&#969;). Because this approach is imaging-based, mechanical characterization is also mapped onto large transmitted-light spatial fields to simultaneously report qualitative changes in 3D tumor size and phenotype. Representative results showing contrasting mechanical response in sub-regions associated with localized invasion-induced matrix degradation as well as system calibration, validation data are presented. Undesirable outcomes from common experimental errors and troubleshooting of these issues are also presented. The 96-well 3D culture plating format implemented in this protocol is conducive to correlation of microrheology measurements with therapeutic screening assays or molecular imaging to gain new insights into impact of treatments or biochemical stimuli on the mechanical microenvironment.

Particle-tracking microrheology can be used to non-destructively quantify and spatially map changes in extracellular matrix mechanical properties in 3D tumor models.

Misurazione longitudinale di matrice extracellulare rigidità in 3D Tumore modelli utilizzando-particelle inseguimento Microrheology

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and ...

Rapid and Low-cost Prototyping of Medical Devices Using 3D Printed Molds for Liquid Injection Molding

Biologically inert elastomers such as silicone are favorable materials for medical device fabrication, but forming and curing these elastomers using traditional liquid injection molding processes can be an expensive process due to tooling and equipment costs. As a result, it has traditionally been impractical to use liquid injection molding for low-cost, rapid prototyping applications. We have devised a method for rapid and low-cost production of liquid elastomer injection molded devices that utilizes fused deposition modeling 3D printers for mold design and a modified desiccator as an injection system. Low costs and rapid turnaround time in this technique lower the barrier to iteratively designing and prototyping complex elastomer devices. Furthermore, CAD models developed in this process can be later adapted for metal mold tooling design, enabling an easy transition to a traditional injection molding process. We have used this technique to manufacture intravaginal probes involving complex geometries, as well as overmolding over metal parts, using tools commonly available within an academic research laboratory. However, this technique can be easily adapted to create liquid injection molded devices for many other applications.

We have devised a method for low-cost and rapid prototyping of liquid elastomer rubber injection molded devices by using fused deposition modeling 3D printers for mold design and a modified desiccator as a liquid injection system.

Rapida e basso costo Prototipazione di dispositivi medici che usano 3D Stampo Stampato per stampaggio ad iniezione Liquid

Biologically inert elastomers such as silicone are favorable materials for medical device fabrication, but forming and curing these elastomers using ...

Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding

The glycoprotein family of collagens represents the main structural proteins in the human body, and are key components of biomaterials used in modern tissue engineering. A technical bottleneck is the deposition of collagen in vitro, as it is notoriously slow, resulting in sub-optimal formation of connective tissue and subsequent tissue cohesion, particularly in skin models. Here, we describe a method which involves the addition of differentially-sized sucrose co-polymers to skin cultures to generate macromolecular crowding (MMC), which results in a dramatic enhancement of collagen deposition. Particularly, dermal fibroblasts deposited a significant amount of collagen I/IV/VII and fibronectin under MMC in comparison to controls.
The protocol also describes a method to decellularize crowded cell layers, exposing significant amounts of extracellular matrix (ECM) which were retained on the culture surface as evidenced by immunocytochemistry. Total matrix mass and distribution pattern was studied using interference reflection microscopy. Interestingly, fibroblasts, keratinocytes and co-cultures produced cell-derived matrices (CDM) of varying composition and morphology. CDM could be used as &#34;bio-scaffolds&#34; for secondary cell seeding, where the current use of coatings or scaffolds, typically from xenogenic animal sources, can be avoided, thus moving towards more clinically relevant applications.
In addition, this protocol describes the application of MMC during the submerged phase of a 3D-organotypic skin co-culture model which was sufficient to enhance ECM deposition in the dermo-epidermal junction (DEJ), in particular, collagen VII, the major component of anchoring fibrils. Electron microscopy confirmed the presence of anchoring fibrils in cultures developed with MMC, as compared to controls. This is significant as anchoring fibrils tether the dermis to the epidermis, hence, having a pre-formed mature DEJ may benefit skin graft recipients in terms of graft stability and overall wound healing. Furthermore, culture time was condensed from 5 weeks to 3 weeks to obtain a mature construct, when using MMC, reducing costs.

Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding

We present a protocol to obtain cell-derived matrices rich in extracellular matrix proteins, using macromolecular crowders (MMC). In addition, we present a protocol which incorporates MMC in 3D organotypic skin co-culture generation, which reduces culture time while maintaining maturity of construct.

Migliorare 2D e 3D della pelle<em&gt; In Vitro</em&gt; modelli con Macromolecole affollamento

The glycoprotein family of collagens represents the main structural proteins in the human body, and are key components of biomaterials used in modern ...

Tunable Hydrogels from Pulmonary Extracellular Matrix for 3D Cell Culture

Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung tissue from porcine, rat, or mouse, the tissue is perfused and submerged in subsequent chemical detergents to remove the cellular debris. Histological comparison of the tissue before and after processing confirms removal of over 95% of double stranded DNA and alpha galactosidase staining suggests the majority of cellular debris is removed. After decellularization, the tissue is lyophilized and then cryomilled into a powder. The matrix powder is digested for 48 hr in an acidic pepsin digestion solution and then neutralized to form the pregel solution. Gelation of the pregel solution can be induced by incubation at 37 &#176;C and can be used immediately following neutralization or stored at 4 &#176;C for up to two weeks. Coatings can be formed using the pregel solution on a non-treated plate for cell attachment. Cells can be suspended in the pregel prior to self-assembly to achieve a 3D culture, plated on the surface of a formed gel from which the cells can migrate through the scaffold, or plated on the coatings. Alterations to the strategy presented can impact gelation temperature, strength, or protein fragment sizes. Beyond hydrogel formation, the hydrogel stiffness may be increased using genipin.

This is a method to create a 3-dimensional cell culture scaffold from pulmonary extracellular matrix. Intact lung is processed into hydrogels that can support the growth of cells in three-dimensions.

Sintonizzabile idrogel da polmonare matrice extracellulare per colture cellulari 3D

Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung ...

Scaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method

Coronary artery disease remains a leading cause of death, affecting millions of Americans. With the lack of autologous vascular grafts available, engineered grafts offer great potential for patient treatment. However, engineered vascular grafts are generally not easily scalable, requiring manufacture of custom molds or polymer tubes in order to customize to different sizes, constituting a time-consuming and costly practice. Human arteries range in lumen diameter from about 2.0-38 mm and in wall thickness from about 0.5-2.5 mm. We have created a method, termed the "Ring Stacking Method," in which variable size rings of tissue of the desired cell type, demonstrated here with vascular smooth muscle cells (SMCs), can be created using guides of center posts to control lumen diameter and outer shells to dictate vessel wall thickness. These tissue rings are then stacked to create a tubular construct, mimicking the natural form of a blood vessel. The vessel length can be tailored by simply stacking the number of rings required to constitute the length needed. With our technique, tissues of tubular forms, similar to a blood vessel, can be readily manufactured in a variety of dimensions and lengths to meet the needs of the clinic and patient.

Scalable engineered blood vessels would improve clinical applicability. Using easily sizable 3D-printed guides, rings of vascular smooth muscle were created and stacked into a tubular form, forming a vascular graft. Grafts can be sized to meet the range of human coronary artery dimensions by simply changing the 3D-printed guide size.

Scaling di Engineered vascolari innesti Usando 3D Stampato Guide e il metodo di impilamento Anello

Coronary artery disease remains a leading cause of death, affecting millions of Americans. With the lack of autologous vascular grafts available, ...

Scaled Anatomical Model Creation of Biomedical Tomographic Imaging Data and Associated Labels for Subsequent Sub-surface Laser Engraving (SSLE) of Glass Crystals

Biomedical imaging modalities like computed tomography (CT) and magnetic resonance (MR) provide excellent platforms for collecting three-dimensional data sets of patient or specimen anatomy in clinical or preclinical settings. However, the use of a virtual, on-screen display limits the ability of these tomographic images to fully convey the anatomical information embedded within. One solution is to interface a biomedical imaging data set with 3D printing technology to generate a physical replica. Here we detail a complementary method to visualize tomographic imaging data with a hand-held model: Sub Surface Laser Engraving (SSLE) of crystal glass. SSLE offers several unique benefits including: the facile ability to include anatomical labels, as well as a scale bar; streamlined multipart assembly of complex structures in one medium; high resolution in the X, Y, and Z planes; and semi-transparent shells for visualization of internal anatomical substructures. Here we demonstrate the process of SSLE with CT data sets derived from pre-clinical and clinical sources. This protocol will serve as a powerful and inexpensive new tool with which to visualize complex anatomical structures for scientists and students in a number of educational and research settings.

Scaled Anatomical Model Creation of Biomedical Tomographic Imaging Data and Associated Labels for Subsequent Sub-surface Laser Engraving SSLE of Glass Crystals

A methodology is described herein for representing anatomical imaging data within crystals. We create scaled three-dimensional models of biomedical imaging data for use in Sub-Surface Laser Engraving (SSLE) of crystal glass. This tool offers a useful complement to computational display or three-dimensionally printed models used within clinical or educational settings.

Scaled Modello anatomico Creazione di Biomedical Imaging tomografica dei dati e associate etichette per la successiva sotto la superficie incisione laser (SSLE) di cristalli di vetro

Biomedical imaging modalities like computed tomography (CT) and magnetic resonance (MR) provide excellent platforms for collecting three-dimensional data ...

Fabrication of Custom Agarose Wells for Cell Seeding and Tissue Ring Self-assembly Using 3D-Printed Molds

Engineered tissues are being used clinically for tissue repair and replacement, and are being developed as tools for drug screening and human disease modeling. Self-assembled tissues offer advantages over scaffold-based tissue engineering, such as enhanced matrix deposition, strength, and function. However, there are few available methods for fabricating 3D tissues without seeding cells on or within a supporting scaffold. Previously, we developed a system for fabricating self-assembled tissue rings by seeding cells into non-adhesive agarose wells. A polydimethylsiloxane (PDMS) negative was first cast in a machined polycarbonate mold, and then agarose was gelled in the PDMS negative to create ring-shaped cell seeding wells. However, the versatility of this approach was limited by the resolution of the tools available for machining the polycarbonate mold. Here, we demonstrate that 3D-printed plastic can be used as an alternative to machined polycarbonate for fabricating PDMS negatives. The 3D-printed mold and revised mold design is simpler to use, inexpensive to produce, and requires significantly less agarose and PDMS per cell seeding well. We have demonstrated that the resulting agarose wells can be used to create self-assembled tissue rings with customized diameters from a variety of different cell types. Rings can then be used for mechanical, functional, and histological analysis, or for fabricating larger and more complex tubular tissues.

This protocol describes a platform for fabricating self-assembled tissue rings in variable sizes using a customized 3D-printed plastic mold. PDMS negatives are cured in the 3D-printed mold; then agarose is cast in the cured PDMS negatives. Cells are seeded into the resulting agarose wells where they aggregate into tissue rings.

Fabbricazione di pozzi di agarosio personalizzato per la semina delle cellule e tessuto anello auto-assemblaggio utilizzando stampi 3D-stampato

Engineered tissues are being used clinically for tissue repair and replacement, and are being developed as tools for drug screening and human disease ...

Targeting Neuronal Fiber Tracts for Deep Brain Stimulation Therapy Using Interactive, Patient-Specific Models

Deep brain stimulation (DBS), which involves insertion of an electrode to deliver stimulation to a localized brain region, is an established therapy for movement disorders and is being applied to a growing number of disorders. Computational modeling has been successfully used to predict the clinical effects of DBS; however, there is a need for novel modeling techniques to keep pace with the growing complexity of DBS devices. These models also need to generate predictions quickly and accurately. The goal of this project is to develop an image processing pipeline to incorporate structural magnetic resonance imaging (MRI) and diffusion weighted imaging (DWI) into an interactive, patient specific model to simulate the effects of DBS. A virtual DBS lead can be placed inside of the patient model, along with active contacts and stimulation settings, where changes in lead position or orientation generate a new finite element mesh and solution of the bioelectric field problem in near real-time, a timespan of approximately 10 seconds. This system also enables the simulation of multiple leads in close proximity to allow for current steering by varying anodes and cathodes on different leads. The techniques presented in this paper reduce the burden of generating and using computational models while providing meaningful feedback about the effects of electrode position, electrode design, and stimulation configurations to researchers or clinicians who may not be modeling experts.

The goal of this project is to develop an interactive, patient-specific modeling pipeline to simulate the effects of deep brain stimulation in near real-time and provide meaningful feedback as to how these devices influence neural activity in the brain.

Tratti della fibra di un neurone di targeting per la terapia di stimolazione cerebrale profonda utilizzando modelli interattivi, specifico per ogni paziente

Deep brain stimulation (DBS), which involves insertion of an electrode to deliver stimulation to a localized brain region, is an established therapy for ...

High-resolution Patterned Biofilm Deposition Using pDawn-Ag43

Spatial structure and patterning play an important role in bacterial biofilms. Here we demonstrate an accessible method for culturing E. coli biofilms into arbitrary spatial patterns at high spatial resolution. The technique uses a genetically encoded optogenetic construct&#8212;pDawn-Ag43&#8212;that couples biofilm formation in E. coli to optical stimulation by blue light. We detail the process for transforming E. coli with&#160;pDawn-Ag43, preparing the required optical set-up, and the protocol for culturing patterned biofilms using pDawn-Ag43 bacteria. Using this protocol, biofilms with a spatial resolution below 25 &#956;m can be patterned on various surfaces and environments, including enclosed chambers, without requiring microfabrication, clean-room facilities, or surface pretreatment. The technique is convenient and appropriate for use in applications that investigate the effect of biofilm structure, providing tunable control over biofilm patterning. More broadly, it also has potential applications in biomaterials, education, and bio-art.

We demonstrate a method for depositing Escherichia coli bacterial biofilms in arbitrary spatial patterns with a high resolution using optical stimulation of a genetically encoded surface-adhesion construct.

Alta risoluzione svevapacifico modellato mediante deposizione di Biofilm-Ag43

Spatial structure and patterning play an important role in bacterial biofilms. Here we demonstrate an accessible method for culturing E. coli biofilms ...