Name: generation of scalable, metallic high-aspect ratio nanocomposites in a biological liquid medium
Uploaded: 2015-07-08T15:00:00
Description: Watch this Scientific Journal Video about Generation of Scalable, Metallic High-Aspect Ratio Nanocomposites in a Biological Liquid Medium at JoVE.com

The authors would like to acknowledge the technical assistance of Alfred Gunasekaran in electron microscopy studies at the Institute of Micromanufacturing at Louisiana Tech University, and Dr. Jim McNamara for assistance with additional microscopy studies. The work described was supported in part by Louisiana board of Regents PKSFI Contract No. LEQSF (2007-12)-ENH-PKSFI-PRS-04 and the James E. Wyche III Endowed Professorship from Louisiana Tech University (to M.D.).

1. תכנון של ניסויים <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לקבוע את עוצמת הקול של nanocomposites הנחושת נחוץ לסינתזה. על בסיס זה, לבחור מספר צלוחיות הקטנות נפח (25 סנטימטר 2), או צלוחיות גדולות יותר כמפורט להלן בהכנת חומרים. </li><li style=";text-align:right;direction:rtl"> לסינתזה ז?...

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-Aspect+Ratio+Bio-Metallic+Nanocomposites+for+Cellular+Interactions.">High-Aspect Ratio Bio-Metallic Nanocomposites for Cellular Interactions.</a> IOP Conference Series: Materials Science and Engineering. 64, 012014 (2014).</li><li>Montes-Burgos, I., 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=Characterisation+of+nanoparticle+size+and+state+prior+to+nanotoxicological+studies.">Characterisation of nanoparticle size and state prior to nanotoxicological studies.</a> Journal of Nanoparticle Research. 12, 47-53 (2010).</li><li>Wiogo, H. 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G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+the+coffee-ring+effect+by+shape-dependent+capillary+interactions.">Suppression of the coffee-ring effect by shape-dependent capillary interactions.</a> Nature. 476, 308-311 (2011).</li><li>Kahler, H., Lloyd Jr, B., Eden, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electron+Microscopic+and+Other+Studies+on+a+Copper&#8211;Cystine+Complex.">Electron Microscopic and Other Studies on a Copper&#8211;Cystine Complex.</a> The Journal of Physical Chemistry. 56, 768-770 (1952).</li><li>Furia, E., Sindona, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complexation+of+L-cystine+with+metal+cations.">Complexation of L-cystine with metal cations.</a> Journal of Chemical &amp; Engineering Data. 55, 2985-2989 (2010).</li><li>Hawkins, C., Perrin, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polynuclear+Complex+Formation.+II.+Copper+(II)+with+Cystine+and+Related+Ligands.">Polynuclear Complex Formation. II. Copper (II) with Cystine and Related Ligands.</a> Inorganic Chemistry. 2, 843-849 (1963).</li><li>Hallman, P., Perrin, D., Watt, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+computed+distribution+of+copper+(II)+and+zinc+(II)+ions+among+seventeen+amino+acids+present+in+human+blood+plasma.">The computed distribution of copper (II) and zinc (II) ions among seventeen amino acids present in human blood plasma.</a> Biochem. J. 121, 549-555 (1971).</li><li>Jensen, L. S., Maurice, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+sulfur+amino+acids+on+copper+toxicity+in+chicks.">Influence of sulfur amino acids on copper toxicity in chicks.</a> The Journal of nutrition. 109, 91-97 (1979).</li><li>Lee, Y., Choi, J. R., Lee, K. J., Stott, N. 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תוך הערכת השפעות רעילות פוטנציאליות של ננו כולל CNPs, זה היה ציין כי בטווח הארוך, CNPs הפכו מהפצת חלקיקים בתחילה יותר מפוזרת לצורה גדולה יותר, מצטברת (איור 2). במקרים מסוימים, תצורות מצטברות מאוד אלה שיוצרו בצלחת תרבית תאים, בתנאים ביולוגיים, יצרו תחזיות ליניארי ב?...

Copper is a highly reactive metal that in the biological world is essential in some enzyme functions 1,2, but in higher concentrations is potently toxic including in the nanoparticulate form 3,4. Concern over copper toxicity has become more relevant as CNPs and other copper-based nanomaterials are utilized, due to the increased surface area/mass for nanostructures. Thus, even a small mass of copper, in nanoparticle form, could cause local toxicity due to its ability to penetrate the cell and be broken down into reactive forms. Some biological species can complex with and chelate metal ions, and even incorporate them into biological structures as has been described in marine mussels 5. In studying the potential toxic effects of nanomaterials 4, it was discovered that over time, and under biological conditions used for typical cell culturing (37 &#176;C and 5% CO2), stable copper biocomposites could be formed with a high-aspect ratio (linear) structure.
By a process of elimination, the initial discovery of these linear biocomposites, which occurred in complete cell culture media, was simplified to a defined protocol of essential elements needed for the biocomposites to self-assemble. Self-assembly of two types of highly linear biocomposites was discovered to be possible with two starting metal components: 1) CNPs and 2) copper sulfate, with the common biological component being cystine. Although more complex, so called &#8220;urchin&#8221; and &#8220;nanoflower&#8221; type copper-containing structures with nanoscale and microscale features have been previously reported, these were produced under non-biological conditions, such as temperatures of 100 &#176;C or greater 6-8. To our knowledge, synthesis of individual, linear copper-containing nanostructures that are scalable in liquid phase under biological conditions has not been previously described. 
One of the starting materials utilized for synthesis of nanocomposites, namely CNPs, has been reported previously to be very toxic to cells 4. It has recently been reported that after the nanocomposites are formed, these structures are less toxic on a per mass basis than the starting NPs 9. Thus, the synthesis described here may be derived from a biological and biochemical reaction that has utility in stabilizing reactive copper species, both in the sense of transforming the NP form into larger structures and in producing composites less toxic to cells. 
In contrast to many other nanomaterial forms which are known to aggregate or clump upon interaction with biological liquid media 10,11, once formed, the highly linear composites described here avoid aggregation, possibly due to a redistribution of charge which has been previously reported 9. As detailed in the current work, this avoidance of aggregation is convenient for the purposes of working with the structures once formed for at least 3 reasons: 1) composite structures once formed may be concentrated using centrifugation and then easily dispersed again using vortex mixing; 2) formed structures can be decreased in average size by sonication for different periods of time; and 3) the formed linear structures may provide an additional tool for avoiding the recently described &#8220;coffee ring effect&#8221; 12 and thus provide a dopant for creating more evenly distributed coatings of materials, especially those containing spherical particulates.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Mini Vortexer</td>
			<td>VWR (https://us.vwr.com)</td>
			<td>58816-121</td>
			<td></td>
		</tr>
		<tr>
			<td>CO2 Incubator Model # 2425-2</td>
			<td>VWR (https://us.vwr.com)</td>
			<td>Contact vendor</td>
			<td>Current model calalog # 98000-360</td>
		</tr>
		<tr>
			<td>Eppendorf Centrifuge (Refrigerated Microcentrifuge)</td>
			<td>Labnet (http://labnetinternational.com/)</td>
			<td>C2500-R</td>
			<td>Model Prism R</td>
		</tr>
		<tr>
			<td>Cell Culture Centrifuge Model Z323K</td>
			<td>Labnet (http://labnetinternational.com/)</td>
			<td>Contact vendor</td>
			<td>Current model Z206A catalog # C0206-A</td>
		</tr>
		<tr>
			<td>Sonicator (Ultrasonic Cleaner)</td>
			<td>Branson Ultrasonics Corporation (http://www.bransonic.com/)</td>
			<td>1510R-MTH</td>
			<td></td>
		</tr>
		<tr>
			<td>Balance</td>
			<td>Sartorius (http://dataweigh.com)</td>
			<td></td>
			<td>Model CP225D similar model CPA225D</td>
		</tr>
		<tr>
			<td>Olympus IX51 Inverted Light Microscope</td>
			<td>Olympus (http://olympusamerica.com</td>
			<td>Contact vendor</td>
			<td></td>
		</tr>
		<tr>
			<td>Olympus DP71 microscope digital camera</td>
			<td>Olympus (http://olympusamerica.com</td>
			<td>Contact vendor</td>
			<td></td>
		</tr>
		<tr>
			<td>external power supply unit- white light for Olympus microscope</td>
			<td>Olympus (http://olympusamerica.com</td>
			<td>TH4-100</td>
			<td></td>
		</tr>
		<tr>
			<td>10x, 20, and 40x microscope objectives</td>
			<td>Olympus (http://olympusamerica.com</td>
			<td>Contact vendor</td>
			<td></td>
		</tr>
		<tr>
			<td>Scanning Electron Microscope</td>
			<td>Hitachi (http://hitachi-hitec.com/global/em/sem/sem_index.html)</td>
			<td>model S-4800</td>
			<td></td>
		</tr>
		<tr>
			<td>Transmission Electron Microscope</td>
			<td>Zeiss (http://zeiss.com/microscopy/en_de/products.html)</td>
			<td>model Libra 120</td>
			<td></td>
		</tr>
		<tr>
			<td>Table Top Work Station Unidirectional Flow Clean Bench</td>
			<td>Envirco (http://envirco-hvac.com)</td>
			<td>model PNG62675</td>
			<td>Used for sterile cell culture technique</td>
		</tr>
	</tbody>
</table>

generation of scalable, metallic high-aspect ratio nanocomposites in a biological liquid medium

המטרה של פרוטוקול זה היא לתאר את הסינתזה של שני biocomposites רומן עם מבני יחס גבוה היבט. Biocomposites מורכב מנחושת וציסטין, גם עם חלקיקי נחושת (CNPs) או נחושת גופרתית תורם הרכיב המתכתי. הסינתזה מתבצעת בנוזל בתנאים ביולוגיים (37 מעלות צלזיוס) וצורת מרוכבים עצמיים התאספו לאחר 24 שעות. ברגע שנוצר, חומרים מרוכבים אלה הם מאוד יציבים בשתי תקשורת הנוזלית ובצורה יבשה. חומרים מרוכבים בקנה מידה מננו למייקרו טווח אורך, ומכמה מיקרונים עד 25 ננומטר בקוטר. מיקרוסקופ אלקטרונים סורקים פליטת שדה עם ספקטרוסקופיה רנטגן נפיצה אנרגיה (EDX) הוכיח כי הגופרית הייתה נוכחת במבנים ליניארי נגזר-NP, בזמן שהוא נעדר מחומר CNP מתחיל, ובכך אישר ציסטין כמקור של גופרית בnanocomposites הסופי . במהלך סינתזה של ננו ומיקרו-חומרים מרוכבים ליניארי אלה, במגוון הרחב של אורכים של structures נוצר בכלי הסינתזה. Sonication של התערובת הנוזלית לאחר הסינתזה הודגם לסייע בשליטה בגודל ממוצע של המבנים על ידי צמצום האורך הממוצע עם זמן מוגבר של sonication. מאז המבנים נוצרו הם מאוד יציבים, לא מצבר, ונוצרים בשלב נוזלי, צנטריפוגה עשויה לשמש גם כדי לסייע בריכוז והפרדת חומרים מרוכבים נוצרו.

איור 1 מציג תרשים זרימה סכמטי של הצעדים כדי ליצור סינתזת biocomposites יניארי המתואר בעבודה זו. CNPs או נחושת גופרתית כמתחיל חומרים בשילוב עם מים סטריליים כדי ליצור פתרון 2 מ&quot;ג / מיליליטר, פתרון זה הוא מעורב וsonicated לספק אפילו תערובת, ופתרון נחושת זה הוא מעורב אז ביחס ...

Watch this Scientific Journal Video about Generation of Scalable, Metallic High-Aspect Ratio Nanocomposites in a Biological Liquid Medium at JoVE.com

Generation of Scalable, Metallic High-Aspect Ratio Nanocomposites in a Biological Liquid Medium

כאן אנו מציגים פרוטוקול לסנתז רומן, biocomposites יחס גבוה היבט בתנאים ביולוגיים ובתקשורת נוזלית. Biocomposites קנה מידה מננומטרים למיקרומטר בקוטר ואורך, בהתאמה. חלקיקי נחושת (CNPs) ונחושת גופרתית בשילוב עם ציסטין הם המרכיבים העיקריים לסינתזה.

דור של, Nanocomposites יחס Scalable מתכתי גבוהה היבט בביולוגי נוזלי בינוני

The goal of this protocol is to describe the synthesis of two novel biocomposites with high-aspect ratio structures. The biocomposites consist of copper ...

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