Gli autori vorrebbero riconoscere la nostra fonte di finanziamento NSERC 20/20 di rete per lo sviluppo della avanzati oftalmici materiali.

Tutti gli esperimenti sono stati eseguiti in conformità e il rispetto di tutti gli orientamenti pertinenti delineate dalla University of Waterloo del comitato etico ricerca sugli animali. Gli occhi bovini sono generosamente donati da un mattatoio locale. 1. Modello Eye <ol><li> Progettazione e produzione di stampi 13 <ol><li> Progettare modelli occhio secondo le dimensioni medie fisiologiche di occhi umani adulti. 13 </li><li> Lasciare uno spazio di 250 micron tra il bulbo oculare ed i pezzi palpebra del modello dell&#39;occhio. Progettare i rispettivi stampi usando software per computer-aided design (CAD). </li><li> Creare un nuovo file o di un file .cad .sldprt con AutoCAD o SolidWorks. Creazione di modelli 3D del globo oculare / palpebra umana. Creare stampi dei modelli e salvare gli stampi come file .stl. </li><li> Importare file .stl in software per la stampante 3D (ad esempio, per makeware replicator2). Specificare i parametri di stampa (posizione, essenzialità, la scala, l&#39;orientamento, la scorrevolezza, ecc </em&gt;) 13. </li><li> Salvare il file come file di G-code per stampanti 3D a leggere. Selezionare i materiali come il PLA (acido polilattico), ABS (acrilonitrile butadiene stirene), PC (policarbonato), o una loro combinazione, per stampare gli stampi 13. </li><li> Installare filamento desiderato del materiale di scelta. Importare il file G-code nella stampante 3D da leggere. Stampa lo stampo. NOTA: In alternativa, produrre stampi oculari utilizzando un computer controllato numerico (CNC) della macchina, se una superficie più liscia sul modello dell&#39;occhio è desiderato. Per la produzione di stampi CNC, materiali per stampi non si limitano più alla plastica termici, ma si estendono a metallo, ceramica e polimeri resistivi, come politetrafluoroetilene. </li><li> Aprire l&#39;interfaccia software CNC che è collegato ad un trapano taglio. Costruire stampi 3D in base alla parte anteriore, superiore, laterale e viste prospettiche degli stampi modello bulbo oculare / palpebra precedentemente costruite in interfaccia software di controllo. Selezionare i parametri appropriati per lalavorazione (formato bit, materiale del substrato, spessore del materiale) e procedere al taglio dello stampo. </li></ol></li><li> Sintesi di Oculari Uso PDMS <ol><li> Utilizzando una siringa, misurare 10 ml volume di PDMS (polidimetilsilossano) base e riempire in una provetta da centrifuga 15-50 ml. Aggiungere 10% w / v della soluzione di elastomero in peso totale di PDMS. Con una bacchetta, mescolare bene le soluzioni. </li><li> Versare la soluzione PDMS nel bulbo oculare e la palpebra stampi. Lasciare le PDMS a stabilirsi a RT O / N (o per almeno 12 ore) per avviare la polimerizzazione e per consentire bolle per sciogliere fuori del polimero. NOTA: assicuratevi che non vi siano bolle rimaste nel PDMS che potrebbe salire o espandere. </li><li> Successivamente, mettere gli stampi in forno a 75 ° C (167 ° F) per 1 ora o 150 ° C (302 ° F) per 5 min. Per un gel morbido, lasciare che i PDMS siedono a temperatura ambiente per almeno 48 ore per polimerizzare completamente. </li><li> Mettere i campioni in un congelatore per alcuni minuti; questo si ridurrà le PDMS e semplificarela rimozione dei campioni dagli stampi. Estrarre i oculari dagli stampi usando una spatola sottile. </li><li> Per la consegna della soluzione nello spazio tra il bulbo oculare e pezzi palpebra, collegare un tubo di politetrafluoroetilene 1/16 &quot;x 1/8&quot; con un connettore tubo accoppiatore uguale gamba 1/16 &quot;e fissarlo al pezzo palpebra al foro tubazione . </li></ol></li><li> Sintesi del bulbo oculare brano nel formato Agarosio NOTA: Il pezzo bulbo oculare può essere sintetizzato utilizzando altri polimeri quali agarosio. La seguente procedura può anche essere modificato per produrre oculari da una varietà di tipi di agar, come PDA (patata destrosio agar) o SDA (Sabouraud Dextrose agar). <ol><li> Per produrre un gel 2% (2 g / 100 ml), misura 2 g di agarosio e mescolare con 100 ml di acqua ultrapura. Portare la soluzione ad ebollizione (100 ° C) in modo che l&#39;agarosio si scioglie completamente. Lasciate che la soluzione raffreddare per 5 min. </li><li> Versare la soluzione nello stampo bulbo oculare e lasciare che la soluzione si raffreddi per 30 minuti a temperatura ambiente. Togliere i pezzi bulbo oculare con una spatola. Conservare l&#39;agar bulbo oculare in un -20 ° C freezer per un uso successivo. Per gli studi di microbiologia, sterilizzare gli stampi bulbo oculare in autoclave e / o UV-irradiazione. </li></ol></li><li> Incorporazione di bovino Cornea su PDMS Eyeball NOTA:. Questo protocollo è stato adattato da Parekh et al 14 <ol><li> Eseguire la dissezione e incorporazione delle cornee bovini in condizioni sterili sotto una cappa a flusso laminare. Acquisire gli occhi e sezionare lo stesso giorno. </li><li> Ruotare la cappa a flusso su per 10 minuti prima dell&#39;uso e sanificare con il 70% di alcol etanolo. Assicurarsi che tutti i materiali e gli strumenti sono sterili in autoclave a 273 ° F / 133 ° C per 45 minuti, e posizionati non meno di 4 pollici dall&#39;ingresso cappa a flusso. </li><li> Immergere l&#39;occhio bovina in un becher contenente una soluzione di iodio-povidone diluito per 2 min. Lavare l&#39;occhio in un bicchiere contenente tampone fosfato (PBS) pH 7.4. Uso di pinze posto delicatamente l&#39;occhio su una capsula di Petri di vetro, corneale a faccia in su. </li><li> Rimuovere il muscolo e tessuto adiposo in eccesso tagliando in corrispondenza dei punti di fissaggio sclerali con le forbici fine scollamento. Smaltire il tessuto in eccesso in un bicchiere sterile designato per rifiuti di origine animale. </li><li> Utilizzando micro-forbici, rimuovere la congiuntiva dall&#39;occhio. Avvolgere l&#39;occhio con garza sterile, mantenendo una distanza di almeno 1 cm dal limbo. </li><li> Usando un bisturi, incidere la sclera circa 2 mm dalla regione limbus e superficialmente in modo da evitare la penetrazione della coroide sottostante e corpo vitreo. estendere con attenzione l&#39;incisione di 360 ° con un bisturi o forbici dissezione senza deformare la cornea dalla sua curvatura naturale. </li><li> Con pinza sottile, rimuovere la cornea dall&#39;occhio. Utilizzando pinze, rimuovere con attenzione qualsiasi tessuto uveale aderente e risciacquare cornea con PBS. </li><li> Conservare la cornea a 31ºC in un contenitore sterile con la culturamedio (come ad esempio Medium 199) contenente 3% siero fetale bovino per mantenere l&#39;umidità dei tessuti e nutrimento delle cellule. </li><li> Prima della sperimentazione, riposare la cornea asportata sul bulbo oculare PDMS, e fissare i due pezzi insieme con una clip-on specializzato. </li></ol></li></ol> 2. Blink-piattaforma <ol><li> Progettazione e produzione della Blink-platform NOTA: Il lampeggio piattaforma si compone di tre parti funzionali: modello dell&#39;occhio (descritto nella sezione 1), sistema di ingranaggi e sistema elettronico. <ol><li> Progettazione e produzione della piattaforma di batter l&#39;utilizzo di CAD e stampa 3D, simile a quella descritta per il modello oculare (punto 1.1). Progettare il sistema di ingranaggi in modo tale che traduce semplice rotazione dei motori nei movimenti laterali e rotativi delle oculari. 15 </li><li> Utilizzando il pignone e ingranaggio, tradurre moto di rotazione di un motore passo-passo nel movimento laterale di un pignone, che è collegato ai pezzi palpebra. </li><li> Usando ilsistema di ingranaggi coniugato, amplificare un moto di rotazione da un motore passo-passo in tre (o più) dei moti rotatori per tre diversi pezzi bulbo oculare. </li><li> Allineare i due sistemi di ingranaggi, uno per la palpebra e uno per bulbo oculare, in modo che la distanza tra i due sono costanti. Assemblare il sistema elettronico con un microcontrollore, scudo motore, e due motori. NOTA: utilizzare due motori passo-passo per fornire i motori di rotazione, che si traduce dal sistema di ingranaggi in un movimento lampeggiante. </li><li> Collegare i due motori passo-passo con un sistema costituito da un scudo motore impilati sul microcontrollore. Collegare e configurare i componenti elettronici a lavorare con i prodotti software open source. </li><li> Programmare il sistema per controllare i parametri del motore quali giri al minuto (RPM), numero di round in avanti, il numero di giri a rovescio, e lo stile di svolta. NOTA: fare riferimento al &quot;file di codice Arduino&quot; supplementare per i dettagli. </li><li> Scaricare il software di sistema dal masito nufacturers &#39;. </li><li> Installare il software e aprirlo. Scrivere il codice per il controllo di motori passo passo nella configurazione desiderata. Collegare il sistema con una sorgente per alimentare il sistema elettronico in modo che i motori si muovono nella maniera desiderata come definito dal ricercatore. NOTA: fare riferimento al &quot;file di codice Arduino&quot; supplementare. </li></ol></li><li> Montaggio con microfluidica (Artificial strappo Solution) <ol><li> Prendere il bulbo oculare sintetizzato e pezzi delle palpebre e li scivolare sui loro corrispondenti clip-on per il modello dell&#39;occhio. Collegare il tubo che si unisce con una siringa e posizionato sulla pompa microfluidica con il pezzo palpebra (sezione 1.2.5). Prova di funzionamento la piattaforma e verificare per il movimento costante. </li><li> Primo il tubo e verificare la presenza di un flusso costante di soluzione di lacrime artificiali (ATS). La ricetta per ATS è stato riportato in precedenza. 16 </li><li> spostare manualmente le parti eye-modello insieme in piano, in modo che il bulbo oculare e l&#39;occhiocoperchio sono in contatto. Impostare la portata della pompa microfluidi ai valori desiderati. Impostare portate fisiologici a 1-1,5 ml / min. 17 </li><li> Avviare la pompa e gli attuatori per iniziare esperimento. Per gli esperimenti di consegna della droga, posizionare la lente a contatto farmaci contenenti sul pezzo bulbo oculare. </li><li> Lasciare che il fluido flusso continuo a goccia a goccia in 12-pozzetti. Ad intervalli di tempo stabiliti desiderati, quantificare la concentrazione dell&#39;analita o di droga utilizzando metodi di rilevamento comuni come la spettroscopia UV-Vis o fluorescenza. 1,4,18 </li><li> Per gli studi che valutano la deposizione di componenti lacrimali su lenti a contatto, posizionare la lente a contatto sul pezzo &quot;bulbo oculare&quot;. Raccogliere il fluido a flusso continuo, che può essere scartata. </li><li> Dopo gli intervalli di tempo desiderati, rimuovere la lente a contatto dal pezzo bulbo oculare e preparare la lente per ulteriore analisi come la microscopia confocale. </li></ol></li></ol>

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	<li>Phan, C. M., Subbaraman, L. N., Jones, L. <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+drug+release+of+natamycin+from+beta-cyclodextrin+and+2-hydroxypropyl-beta-cyclodextrin-functionalized+contact+lens+materials.">In vitro drug release of natamycin from beta-cyclodextrin and 2-hydroxypropyl-beta-cyclodextrin-functionalized contact lens materials.</a> J Biomater Sci Polym Ed. 25, 1907-1919 (2014).</li><li>Peng, C. C., Kim, J., Chauhan, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extended+delivery+of+hydrophilic+drugs+from+silicone-hydrogel+contact+lenses+containing+vitamin+E+diffusion+barriers.">Extended delivery of hydrophilic drugs from silicone-hydrogel contact lenses containing vitamin E diffusion barriers.</a> Biomaterials. 31, 4032-4047 (2010).</li><li>Hui, A., Willcox, M., Jones, L. <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+and+in+vivo+evaluation+of+novel+ciprofloxacin-releasing+silicone+hydrogel+contact+lenses.">In vitro and in vivo evaluation of novel ciprofloxacin-releasing silicone hydrogel contact lenses.</a> Invest Ophthalmol Vis Sci. 55, 4896-4904 (2014).</li><li>Boone, A., Hui, A., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+and+release+of+dexamethasone+phosphate+from+silicone+hydrogel+and+group+I,+II,+and+IV+hydrogel+contact+lenses.">Uptake and release of dexamethasone phosphate from silicone hydrogel and group I, II, and IV hydrogel contact lenses.</a> Eye Contact Lens. 35, 260-267 (2009).</li><li>Lorentz, H., Heynen, M., Trieu, D., Hagedorn, S. J., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+tear+film+components+on+in+vitro+lipid+uptake.">The impact of tear film components on in vitro lipid uptake.</a> Optom Vis Sci. 89, 856-867 (2012).</li><li>Hall, B., Phan, C. M., Subbaraman, L., Jones, L. W., Forrest, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extraction+versus+in+situ+techniques+for+measuring+surface-adsorbed+lysozyme.">Extraction versus in situ techniques for measuring surface-adsorbed lysozyme.</a> Optom Vis Sci. 91, 1062-1070 (2014).</li><li>Mishima, S., Gasset, A., Klyce, S. D., Baum, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+tear+volume+and+tear+flow.">Determination of tear volume and tear flow.</a> Invest Ophthalmol Vis Sci. 5, 264-276 (1966).</li><li>Nichols, J. 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=The+TFOS+international+workshop+on+contact+lens+discomfort:+executive+summary.">The TFOS international workshop on contact lens discomfort: executive summary.</a> Invest Ophthalmol Vis Sci. 54, 7-13 (2013).</li><li>Peng, C. C., Burke, M. T., Carbia, B. E., Plummer, C., Chauhan, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extended+drug+delivery+by+contact+lenses+for+glaucoma+therapy.">Extended drug delivery by contact lenses for glaucoma therapy.</a> J Control Release. 162, 152-158 (2012).</li><li>Faschinger, C., Mossbock, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Continuous+24+h+monitoring+of+changes+in+intraocular+pressure+with+the+wireless+contact+lens+sensor+Triggerfish.+First+results+in+patients.">Continuous 24 h monitoring of changes in intraocular pressure with the wireless contact lens sensor Triggerfish. First results in patients.</a> Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft. 107, 918-922 (2010).</li><li>Shaw, A. J., Davis, B. A., Collins, M. J., Carney, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+to+measure+eyelid+pressure+using+piezoresistive+sensors.">A technique to measure eyelid pressure using piezoresistive sensors.</a> IEEE transactions on bio-medical engineering. 56, 2512-2517 (2009).</li><li>Liao, Y. T., Yao, H. F., Lingley, A., Parviz, B., Otis, B. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+3-mu+W+CMOS+glucose+sensor+for+wireless+contact-lens+tear+glucose+monitoring.">A 3-mu W CMOS glucose sensor for wireless contact-lens tear glucose monitoring.</a> Ieee J Solid-St Circ. 47, 335-344 (2012).</li><li>Coster, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Cornea. , (2002).</li><li>Parekh, 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=A+simplified+technique+for+in+situ+excision+of+cornea+and+evisceration+of+retinal+tissue+from+human+ocular+globe.">A simplified technique for in situ excision of cornea and evisceration of retinal tissue from human ocular globe.</a> Journal of visualized experiments : JoVE. , e3765 (2012).</li><li>Way, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gear+and+pinion.">Gear and pinion.</a> US patent. , (1942).</li><li>Lorentz, 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=Contact+lens+physical+properties+and+lipid+deposition+in+a+novel+characterized+artificial+tear+solution.">Contact lens physical properties and lipid deposition in a novel characterized artificial tear solution.</a> Molecular vision. 17, 3392-3405 (2011).</li><li>Furukawa, R. E., Polse, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+tear+flow+accompanying+aging.">Changes in tear flow accompanying aging.</a> American journal of optometry and physiological optics. 55, 69-74 (1978).</li><li>Bajgrowicz, M., Phan, C. M., Subbaraman, L., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Release+of+ciprofloxacin+and+moxifloxacin+from+daily+disposable+contact+lenses+from+an+in+vitro+eye+model.">Release of ciprofloxacin and moxifloxacin from daily disposable contact lenses from an in vitro eye model.</a> Invest Ophthalmol Vis Sci. , (2015).</li><li>Luensmann, D., Zhang, F., Subbaraman, L., Sheardown, H., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+lysozyme+sorption+to+conventional+and+silicone+hydrogel+contact+lenses+using+confocal+microscopy.">Localization of lysozyme sorption to conventional and silicone hydrogel contact lenses using confocal microscopy.</a> Current eye research. 34, 683-697 (2009).</li><li>Tieppo, A., Pate, K. M., Byrne, M. E. <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+controlled+release+of+an+anti-inflammatory+from+daily+disposable+therapeutic+contact+lenses+under+physiological+ocular+tear+flow.">In vitro controlled release of an anti-inflammatory from daily disposable therapeutic contact lenses under physiological ocular tear flow.</a> Eur J Pharm Biopharm. 81, 170-177 (2012).</li><li>Ali, 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=Zero-order+therapeutic+release+from+imprinted+hydrogel+contact+lenses+within+in+vitro+physiological+ocular+tear+flow.">Zero-order therapeutic release from imprinted hydrogel contact lenses within in vitro physiological ocular tear flow.</a> J Control Release. 124, 154-162 (2007).</li><li>White, C. J., McBride, M. K., Pate, K. M., Tieppo, A., Byrne, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extended+release+of+high+molecular+weight+hydroxypropyl+methylcellulose+from+molecularly+imprinted,+extended+wear+silicone+hydrogel+contact+lenses.">Extended release of high molecular weight hydroxypropyl methylcellulose from molecularly imprinted, extended wear silicone hydrogel contact lenses.</a> Biomaterials. 32, 5698-5705 (2011).</li><li>Kaczmarek, J. C., Tieppo, A., White, C. J., Byrne, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adjusting+biomaterial+composition+to+achieve+controlled+multiple-day+release+of+dexamethasone+from+an+extended-wear+silicone+hydrogel+contact+lens.">Adjusting biomaterial composition to achieve controlled multiple-day release of dexamethasone from an extended-wear silicone hydrogel contact lens.</a> J Biomater Sci Polym Ed. 25, 88-100 (2014).</li><li>Mohammadi, S., Postnikoff, C., Wright, A. M., Gorbet, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+development+of+an+in+vitro+tear+replenishment+system.">Design and development of an in vitro tear replenishment system.</a> Ann Biomed Eng. 42, 1923-1931 (2014).</li><li>Lorentz, H., Heynen, M., Khan, W., Trieu, D., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+intermittent+air+exposure+on+lipid+deposition.">The impact of intermittent air exposure on lipid deposition.</a> Optom Vis Sci. 89, 1574-1581 (2012).</li><li>Peng, C. C., Fajardo, N. P., Razunguzwa, T., Radke, C. J. <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+spoilation+of+silicone-hydrogel+soft+contact+lenses+in+a+model-blink+cell.">In vitro spoilation of silicone-hydrogel soft contact lenses in a model-blink cell.</a> Optom Vis Sci. 92, 768-780 (2015).</li><li>Liu, 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=Dissolution+studies+of+poorly+soluble+drug+nanosuspensions+in+non-sink+conditions.">Dissolution studies of poorly soluble drug nanosuspensions in non-sink conditions.</a> AAPS PharmSciTech. 14, 748-756 (2013).</li></ol>

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Ci sono tre fasi critiche all&#39;interno del protocollo che richiedono particolare attenzione: la progettazione e produzione di stampi (punto 1.1), il montaggio piattaforma (sezione 2.2.1-2.2.3), e monitorare il percorso sperimentale (sezione 2.2.4-2.2.7 ). In termini di progettazione e produzione di stampi (sezione 1.1), il pezzo bulbo oculare dovrebbe essere progettato in funzione delle dimensioni di una cornea umana. Tuttavia, può richiedere più prototipi dello stampo prima che un pezzo bulbo oculare può essere c...

Due significative aree di interesse all&#39;interno dell&#39;arena di lenti a contatto (CL) comprendono il disagio e lo sviluppo di applicazioni innovative CL. Chiarire i meccanismi alla base CL disagio è un problema che ha eluso il campo per decenni. 8 Lo sviluppo del romanzo, CL funzionali, come i dispositivi di consegna di droga 1,3,9 e biosensori, 10-12 è un&#39;area di crescente interesse, con i mercati potenziali sostanziali. In entrambi i casi, un sofisticato modello in vitro potrebbe fornire informazioni utili per aiutare con la selezione appropriata materiali per lenti o caratteristiche di progettazione durante la fase di sviluppo. Purtroppo, corrente in modelli in vitro per la valutazione CL e altre applicazioni relative oculari sono relativamente grezzo e sofisticato. Tradizionalmente, studi in vitro CL valutare la deposizione di film lacrimale o la consegna di droga sono eseguite in, grandi fiale di volume statiche contenenti un volume di fluido fisso, che Greasupera tly quantità fisiologiche. Inoltre, questo semplice modello manca il componente flusso lacrimale naturale e il riflesso lampeggiante, entrambi i quali sono definendo fattori dell&#39;ambiente oculare. Lo sviluppo di un fisiologicamente rilevanti &quot;modello&quot; sofisticato, occhio richiederà un approccio multi-disciplinare e richiedono sostanziale validazione vivo. Per queste ragioni, il quadro fondamentale per il nostro modello in vitro occhio è altamente versatile, tale che il modello può essere continuamente migliorata attraverso aggiornamenti e modulazioni futuri. Fino ad oggi, il modello è in grado di simulare volume lacrimale, lacrimazione, usura meccanica e l&#39;esposizione all&#39;aria. Lo scopo è quello di creare un modello in vitro che fornirà risultati significativi, che è predittiva e gratuito per in vivo ed ex vivo osservazioni.

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	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Arduino Uno R3 (Atmega328 - assembled)</td>
			<td style="width:125px;">Adafruit</td>
			<td style="width:451px;">50</td>
			<td style="width:273px;">Board</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Stepper motor</td>
			<td style="width:125px;">Adafruit</td>
			<td style="width:451px;">324</td>
			<td>Motor and Motor shield</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Equal Leg Coupler 1.6mm 1/16&#34;</td>
			<td style="width:125px;">VWR</td>
			<td style="width:451px;">CA11009-280</td>
			<td>50 pcs of tube connector</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Tubing PT/SIL 1/16&#34;x1/8&#34;</td>
			<td style="width:125px;">VWR</td>
			<td style="width:451px;">16211-316</td>
			<td>Case of 50feet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">PDMS</td>
			<td style="width:125px;">Dow Corning</td>
			<td style="width:451px;">Sylgard 184 Solar Cell Encapsulation</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Agarose, Type 1-A, low EEO</td>
			<td style="width:125px;">Sigma-Aldrich</td>
			<td style="width:451px;">A0169-25G</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">PHD UltraTM</td>
			<td style="width:125px;">Harvard Apparatus</td>
			<td style="width:451px;">703006</td>
			<td>MicroFluidic Pump</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Bovine cornea</td>
			<td style="width:125px;">Cargill, Guelph/ON</td>
			<td style="width:451px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Soldidworks</td>
			<td style="width:125px;">Dassault Systemes</td>
			<td style="width:451px;"></td>
			<td>Software</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:281px;">3-D printing</td>
			<td style="width:125px;">University of Waterloo - 3D Print Centre</td>
			<td style="width:451px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Dissection tools</td>
			<td style="width:125px;">Fine Science Tools</td>
			<td style="width:451px;"></td>
			<td>General dissection tools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Medium 199</td>
			<td style="width:125px;">Sigma-Aldrich</td>
			<td style="width:451px;"></td>
			<td>Culture medium storage for cornea</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Fetal bovine serum</td>
			<td style="width:125px;">Thermo Fisher</td>
			<td style="width:451px;"></td>
			<td>Add to culture medium, 3% total volume</td>
		</tr>
	</tbody>
</table>

development of an in vitro ocular platform to test contact lenses

Currently, in vitro evaluations of contact lenses (CLs) for drug delivery are typically performed in large volume vials,1-6 which fail to mimic physiological tear volumes.7 The traditional model also lacks the natural tear flow component and the blinking reflex, both of which are defining factors of the ocular environment. The development of a novel model is described in this study, which consists of a unique 2-piece design, eyeball and eyelid piece, capable of mimicking physiological tear volume. The models are created from 3-D printed molds (Polytetrafluoroethylene or Teflon molds), which can be used to generate eye models from various polymers, such as polydimethylsiloxane (PDMS) and agar. Further modifications to the eye pieces, such as the integration of an explanted human or animal cornea or human corneal construct, will permit for more complex in vitro ocular studies. A commercial microfluidic syringe pump is integrated with the platform to emulate physiological tear secretion. Air exposure and mechanical wear are achieved using two mechanical actuators, of which one moves the eyelid piece laterally, and the other moves the eyeballeyepiece circularly. The model has been used to evaluate CLs for drug delivery and deposition of tear components on CLs.

Gli stampi occhio sintetizzati ottenute dalla officina e dalla stampa 3-D sono riportati nella Figura 1. Questi stampi possono essere utilizzati con una varietà di polimeri, come PDMS e agarosio, per produrre oculari con le proprietà desiderate. L&#39;assemblaggio cenno della piattaforma modello occhio con una pompa a siringa microfluidica è mostrato in Figura 2. La piattaforma simula usura meccanica tramite la rotazione del pezzo bulbo oculare, e l&#...

Watch this Scientific Journal Video about Development of an In Vitro Ocular Platform to Test Contact Lenses at JoVE.com

Development of an In Vitro Ocular Platform to Test Contact Lenses

I modelli attuali in vitro per valutare le lenti a contatto (CLS) e altre applicazioni oculari legate sono molto limitate. La piattaforma oculare presentato simula il flusso fisiologico lacrimale, volume lacrimale, l&#39;esposizione all&#39;aria e l&#39;usura meccanica. Questo sistema è molto versatile e può essere applicato a varie analisi in vitro con CL.

Sviluppo di un<em&gt; In Vitro</em&gt; Oculare piattaforma per testare Lenti a contatto

Currently, in vitro evaluations of contact lenses (CLs) for drug delivery are typically performed in large volume vials,1-6 which fail to mimic ...

development-of-an-in-vitro-ocular-platform-to-test-contact-lenses

Universidad de Cartagena UDEC

Research

JoVE Journal

Bioengineering

Development of an In Vitro Ocular Platform to Test Contact Lenses

10.5K Views.  University of Waterloo.  I modelli attuali in vitro per valutare le lenti a contatto (CLS) e altre applicazioni oculari legate sono molto limitate. La piattaforma oculare presentato simula il flusso fisiologico lacrimale, volume lacrimale, l'esposizione all'aria e l'usura meccanica. Questo sistema è molto versatile e può essere applicato a varie analisi in vitro con CL. 

Video: Development of an In Vitro Ocular Platform to Test Contact Lenses

Tissue Engineering of a Human 3D in vitro Tumor Test System

Cancer is one of the leading causes of death worldwide. Current therapeutic strategies are predominantly developed in 2D culture systems, which inadequately reflect physiological conditions in vivo. Biological 3D matrices provide cells an environment in which cells can self-organize, allowing the study of tissue organization and cell differentiation. Such scaffolds can be seeded with a mixture of different cell types to study direct 3D cell-cell-interactions. To mimic the 3D complexity of cancer tumors, our group has developed a 3D in vitro tumor test system.	
	Our 3D tissue test system models the in vivo situation of malignant peripheral nerve sheath tumors (MPNSTs), which we established with our decellularized porcine jejunal segment derived biological vascularized scaffold (BioVaSc). In our model, we reseeded a modified BioVaSc matrix with primary fibroblasts, microvascular endothelial cells (mvECs) and the S462 tumor cell line. For static culture, the vascular structure of the BioVaSc is removed and the remaining scaffold is cut open on one side (Small Intestinal Submucosa SIS-Muc). The resulting matrix is then fixed between two metal rings (cell crowns).	
	Another option is to culture the cell-seeded SIS-Muc in a flow bioreactor system that exposes the cells to shear stress. Here, the bioreactor is connected to a peristaltic pump in a self-constructed incubator. A computer regulates the arterial oxygen and nutrient supply via parameters such as blood pressure, temperature, and flow rate. This setup allows for a dynamic culture with either pressure-regulated pulsatile or constant flow.	
	In this study, we could successfully establish both a static and dynamic 3D culture system for MPNSTs. The ability to model cancer tumors in a more natural 3D environment will enable the discovery, testing, and validation of future pharmaceuticals in a human-like model.

Tissue Engineering of a Human 3D in vitro Tumor Test System

Methods to create human 3D tumor tissues as test systems are described. These technologies are based on a decellularized Biological Vascularized Scaffold (BioVaSc), primary human cells and a tumor cell line, which can be cultured under static as well as under dynamic conditions in a flow bioreactor.

Ingegneria dei tessuti di un Umano 3D<em&gt; In vitro</em&gt; Sistema Tumore test

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

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Development of Amelogenin-chitosan Hydrogel for In Vitro Enamel Regrowth with a Dense Interface

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or erosion. Amelogenin has been proven to be a critical protein for controlling the organized growth of apatite crystals. In this paper, we present a detailed protocol for superficial enamel reconstruction by using a novel amelogenin-chitosan hydrogel. Compared to other conventional treatments, such as topical fluoride and mouthwash, this method not only has the potential to prevent the development of dental caries but also promotes significant and durable enamel restoration. The organized enamel-like microstructure regulated by amelogenin assemblies can significantly improve the mechanical properties of etched enamel, while the dense enamel-restoration interface formed by an in situ regrowth of apatite crystals can improve the effectiveness and durability of restorations. Furthermore, chitosan hydrogel is easy to use and can suppress bacterial infection, which is the major risk factor for the occurrence of dental caries. Therefore, this biocompatible and biodegradable amelogenin-chitosan hydrogel shows promise as a biomaterial for the prevention, restoration, and treatment of defective enamel.

Development of Amelogenin-chitosan Hydrogel for In Vitro Enamel Regrowth with a Dense Interface

In this article, we describe a protocol for fabricating an amelogenin-chitosan hydrogel for superficial enamel reconstruction. Organized in situ growth of apatite crystals in the hydrogel formed a dense enamel-restoration interface, which will improve the effectiveness and durability of restorations.

Sviluppo di amelogenina-chitosano Hydrogel per<em&gt; In Vitro</em&gt; Smalto ricrescita con una interfaccia Dense

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or ...

Eye Irritation Test (EIT) for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial (RhCE) Tissue Model

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and accurate assessment of ocular toxicity in man are needed. To address this need, we have developed an eye irritation test (EIT) which utilizes a three dimensional reconstructed human cornea-like epithelial (RhCE) tissue model that is based on normal human cells. The EIT is able to separate ocular&#160;irritants and corrosives (GHS Categories 1 and 2 combined) and those that do not require labeling (GHS No Category). The test utilizes two separate protocols, one designed for liquid chemicals and a second, similar protocol for solid test articles. The EIT prediction model uses a single exposure period (30 min for liquids, 6 hr for solids) and a single tissue viability cut-off (60.0% as determined by the MTT assay). Based on the results for 83 chemicals (44 liquids and 39 solids) EIT achieved 95.5/68.2/ and 81.8% sensitivity/specificity and accuracy (SS&#38;A) for liquids, 100.0/68.4/ and 84.6% SS&#38;A for solids, and 97.6/68.3/ and 83.1% for overall SS&#38;A. The EIT will contribute significantly to classifying the ocular irritation potential of a wide range of liquid and solid chemicals without the use of animals to meet regulatory testing requirements. The EpiOcular EIT method was implemented in 2015 into the OECD Test Guidelines as TG 492.

Eye Irritation Test EIT for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial RhCE Tissue Model

We have developed an eye irritation test which utilizes a three dimensional reconstructed human cornea-like epithelial (RhCE) tissue model. The test is able to discriminate between ocular irritant and corrosive materials (GHS Categories 1 and 2 combined) and those that do not require labeling (GHS No Category).

Oculare Test (EIT) per identificazione del pericolo di sostanze chimiche irritanti oculari utilizzando Ricostruito umana Cornea come epiteliale (RHCE) Tessuto Modello

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and ...

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling

Vascular disease is a common cause of death within the United States. Herein, we present a method to examine the contribution of flow dynamics towards vascular disease pathologies. Unhealthy arteries often present with wall stiffening, scarring, or partial stenosis which may all affect fluid flow rates, and the magnitude of pulsatile flow, or pulsatility index. Replication of various flow conditions is the result of tuning a flow pressure damping chamber downstream of a blood pump. Introduction of air within a closed flow system allows for a compressible medium to absorb pulsatile pressure from the pump, and therefore vary the pulsatility index. The method described herein is simply reproduced, with highly controllable input, and easily measurable results. Some limitations are recreation of the complex physiological pulse waveform, which is only approximated by the system. Endothelial cells, smooth muscle cells, and fibroblasts are affected by the blood flow through the artery. The dynamic component of blood flow is determined by the cardiac output and arterial wall compliance. Vascular cell mechano-transduction of flow dynamics may trigger cytokine release and cross-talk between cell types within the artery. Co-culture of vascular cells is a more accurate picture reflecting cell-cell interaction on the blood vessel wall and vascular response to mechanical signaling. Contribution of flow dynamics, including the cell response to the dynamic and mean (or steady) components of flow, is therefore an important metric in determining disease pathology and treatment efficacy. Through introducing an in vitro co-culture model and pressure damping downstream of blood pump which produces simulated cardiac output, various arterial disease pathologies may be investigated.

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling

This protocol replicates physiological or pathological blood flow in vitro to aid in determining cell response in disease pathologies. By introducing a pressure damping chamber downstream of a blood pump, blood flow across the vasculature can be recapitulated and imposed on a monolayer of vascular endothelium or a mimetic co-culture.

Modello in vitro di fisiologiche e patologiche flusso sanguigno con applicazione di indagini di rimodellamento vascolare cellulare

Vascular disease is a common cause of death within the United States. Herein, we present a method to examine the contribution of flow dynamics towards ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices. Whole organ culture of the FSU preserves the native extracellular matrix, cell differentiation phenotypes, and cellular-matrix interactions. Thus, organ culture techniques are particularly useful for investigating the complex biological mechanisms of the IVD. Here, we describe a high-throughput approach for culturing whole lumbar mouse FSUs that provides an ideal platform for studying disease mechanisms and therapies for the IVD. Furthermore, we describe several applications that utilize this organ culture method to conduct further studies including contrast-enhanced microCT imaging and three-dimensional high-resolution finite element modeling of the IVD.

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Whole organ culture of the intervertebral disc (IVD) preserves the native extracellular matrix, cell phenotypes, and cellular-matrix interactions. Here we describe an IVD culture system using mouse lumbar and caudal IVDs in their functional spinal units and several applications utilizing this system.

Un<em&gt; In Vitro</em&gt; Organ Culture Modello del disco intervertebrale Murine

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

Assembly and Tracking of Microbial Community Development within a Microwell Array Platform

The development of microbial communities depends on a combination of complex deterministic and stochastic factors that can dramatically alter the spatial distribution and activities of community members. We have developed a microwell array platform that can be used to rapidly assemble and track thousands of bacterial communities in parallel. This protocol highlights the utility of the platform and describes its use for optically monitoring the development of simple, two-member communities within an ensemble of arrays within the platform. This demonstration uses two mutants of Pseudomonas aeruginosa, part of a series of mutants developed to study Type VI secretion pathogenicity. Chromosomal inserts of either mCherry or GFP genes facilitate the constitutive expression of fluorescent proteins with distinct emission wavelengths that can be used to monitor community member abundance and location within each microwell. This protocol describes a detailed method for assembling mixtures of bacteria into the wells of the array and using time-lapse fluorescence imaging and quantitative image analysis to measure the relative growth of each member population over time. The seeding and assembly of the microwell platform, the imaging procedures necessary for the quantitative analysis of microbial communities within the array, and the methods that can be used to reveal interactions between microbial species area all discussed.

The development of microbial communities depends on a combination of factors, including environmental architecture, member abundance, traits, and interactions. This protocol describes a synthetic, microfabricated environment for the simultaneous tracking of thousands of communities contained in femtoliter wells, where key factors such as niche size and confinement can be approximated.

Assemblea e monitoraggio dello sviluppo microbico della comunità all&#39;interno di una piattaforma di array Microwell

The development of microbial communities depends on a combination of complex deterministic and stochastic factors that can dramatically alter the spatial ...

Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, significantly affecting the rate of human mortality and morbidity. Conventional methods for the detection of V. parahaemolyticus such as culture-based methods, immunological assays, and molecular-based methods require complicated sample handling and are time-consuming, tedious, and costly. Recently, biosensors have proven to be a promising and comprehensive detection method with the advantages of fast detection, cost-effectiveness, and practicality. This research focuses on developing a rapid method of detecting V. parahaemolyticus with high selectivity and sensitivity using the principles of DNA hybridization. In the work, characterization of synthesized polylactic acid-stabilized gold nanoparticles (PLA-AuNPs) was achieved using X-ray Diffraction (XRD), Ultraviolet-visible Spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Field-emission Scanning Electron Microscopy (FESEM), and Cyclic Voltammetry (CV). We also carried out further testing of stability, sensitivity, and reproducibility of the PLA-AuNPs. We found that the PLA-AuNPs formed a sound structure of stabilized nanoparticles in aqueous solution. We also observed that the sensitivity improved as a result of the smaller charge transfer resistance (Rct) value and an increase of active surface area (0.41 cm2). The development of our DNA biosensor was based on modification of a screen-printed carbon electrode (SPCE) with PLA-AuNPs and using methylene blue (MB) as the redox indicator. We assessed the immobilization and hybridization events by differential pulse voltammetry (DPV). We found that complementary, non-complementary, and mismatched oligonucleotides were specifically distinguished by the fabricated biosensor. It also showed reliably sensitive detection in cross-reactivity studies against various food-borne pathogens and in the identification of V. parahaemolyticus in fresh cockles.

A protocol for the development of an electrochemical DNA biosensor comprising a polylactic acid-stabilized, gold nanoparticles-modified, screen-printed carbon electrode to detect Vibrio parahaemolyticus is presented.

Sviluppo di un biosensore elettrochimico del DNA per rilevare un patogeni di origine alimentare

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, ...

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of cellular heterogeneity in biological systems. However, single-cell sequencing of large populations has been hampered by limitations in processing genomes for sequencing. In this paper, we describe a method for single-cell genome sequencing (SiC-seq) which uses droplet microfluidics to isolate, amplify, and barcode the genomes of single cells. Cell encapsulation in microgels allows the compartmentalized purification and tagmentation of DNA, while a microfluidic merger efficiently pairs each genome with a unique single-cell oligonucleotide barcode, allowing &gt;50,000 single cells to be sequenced per run. The sequencing data is demultiplexed by barcode, generating groups of reads originating from single cells. As a high-throughput and low-bias method of single-cell sequencing, SiC-seq will enable a broader range of genomic studies targeted at diverse cell populations.

Single-cell sequencing reveals genotypic heterogeneity in biological systems, but current technologies lack the throughput necessary for the deep profiling of community composition and function. Here, we describe a microfluidic workflow for sequencing &gt;50,000 single-cell genomes from diverse cell populations.

Una piattaforma di Microfluidic ad altissima velocità di trasmissione per il sequenziamento del genoma di cella singola

Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of ...

Rodent Behavioral Testing to Assess Functional Deficits Caused by Microelectrode Implantation in the Rat Motor Cortex

Medical devices implanted in the brain hold tremendous potential. As part of a Brain Machine Interface (BMI) system, intracortical microelectrodes demonstrate the ability to record action potentials from individual or small groups of neurons. Such recorded signals have successfully been used to allow patients to interface with or control&#160;computers, robotic limbs, and their own limbs. However, previous animal studies have shown that a microelectrode implantation in the brain not only damages the surrounding tissue but can also result in functional deficits. Here, we discuss a series of behavioral tests to quantify potential motor impairments following the implantation of intracortical microelectrodes into the motor cortex of a rat. The methods for open field grid, ladder crossing, and grip strength testing provide valuable information regarding the potential complications resulting from a microelectrode implantation. The results of the behavioral testing are correlated with endpoint histology, providing additional information on the pathological outcomes and impacts of this procedure on the adjacent tissue.

We have shown that a microelectrode implantation in the motor cortex of rats causes immediate and lasting motor deficits. The methods proposed herein outline a microelectrode implantation surgery and three rodent behavioral tasks to elucidate potential changes in the fine or gross motor function due to implantation-caused damage to the motor cortex.

Roditore comportamentali test per valutare deficit funzionali causati da microelettrodo impianto nella corteccia di motore del ratto

Medical devices implanted in the brain hold tremendous potential. As part of a Brain Machine Interface (BMI) system, intracortical microelectrodes ...

Advanced 3D Liver Models for In vitro Genotoxicity Testing Following Long-Term Nanomaterial Exposure

Due to the rapid development and implementation of a diverse array of engineered nanomaterials (ENM), exposure to ENM is inevitable and the development of robust, predictive in vitro test systems is essential. Hepatic toxicology is key when considering ENM exposure, as the liver serves a vital role in metabolic homeostasis and detoxification as well as being a major site of ENM accumulation post exposure. Based upon this and the accepted understanding that 2D hepatocyte models do not accurately mimic the complexities of intricate multi-cellular interactions and metabolic activity observed in vivo, there is a greater focus on the development of physiologically relevant 3D liver models tailored for ENM hazard assessment purposes in vitro. In line with the principles of the 3Rs to replace, reduce and refine animal experimentation, a 3D HepG2 cell-line based liver model has been developed, which is a user friendly, cost effective system that can support both extended and repeated ENM exposure regimes (&#8804;14 days). These spheroid models (&#8805;500 &#181;m in diameter) retain their proliferative capacity (i.e., dividing cell models) allowing them to be coupled with the &#8216;gold standard&#8217; micronucleus assay to effectively assess genotoxicity in vitro. Their ability to report on a range of toxicological endpoints (e.g., liver function, (pro-)inflammatory response, cytotoxicity and genotoxicity) has been characterized using several ENMs across both acute (24 h) and long-term (120 h) exposure regimes. This 3D in vitro hepatic model has the capacity to be utilized for evaluating more realistic ENM exposures, thereby providing a future in vitro approach to better support ENM hazard assessment in a routine and easily accessible manner.

This procedure was established to be used for developing advanced 3D hepatic cultures in vitro, which can provide a more physiologically relevant assessment of the genotoxic hazards associated with nanomaterial exposures over both an acute or long-term, repeated dose regimes.

Modelli avanzati di fegato 3D per test di genotossicità in vitro a seguito dell'esposizione a lungo termine ai nanomateriali

Due to the rapid development and implementation of a diverse array of engineered nanomaterials (ENM), exposure to ENM is inevitable and the development of ...

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Ingegneria dei tessuti di un Umano 3D

Sviluppo di amelogenina-chitosano Hydrogel per