Name: magnetic resonance elastography methodology for the evaluation of tissue engineered construct growth
Uploaded: 2012-02-09T13:00:00
Description: Watch this Scientific Journal Video about Magnetic Resonance Elastography Methodology for the Evaluation of Tissue Engineered Construct Growth at JoVE.com

Denne forskning blev delvist støttet af NIH RO3-EB007299-02 og NSF EPSCoR First Award.

1. Tissue Construct Forberedelse Vævet konstruktion fremstillingsprocessen består af tre faser: Udvidelsen af ​​cellepopulationen, podning af celler på et biomateriale stillads, og differentiering ved anvendelse af kemiske signalmolekyler. Proceduren for konstruktionen præparat baseret på metoder, der foretages af Dennis et al., Hong et al. Og Marion og Mao 8,9,10. <ol><li> Efter dyrkning og udvidelse af den cellelinie, frø de humane mesenchymale stam...

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	<li>Xu, H., Othman, S. F., Magin, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+tissue+engineering+using+magnetic+resonance+imaging.">Monitoring tissue engineering using magnetic resonance imaging.</a> J. Biosci. Bioeng. 106, 515-527 (2008).</li><li>Xu, H., Othman, S. F., Hong, L., Peptan, I. A., Magin, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+resonance+microscopy+for+monitoring+osteogenesis+in+tissue-engineered+construct+in+vitro.">Magnetic resonance microscopy for monitoring osteogenesis in tissue-engineered construct in vitro.</a> Phys. Med. Biol. 51, 719-732 (2006).</li><li>Othman, S. F., Xu, H., Royston, T. J., Magin, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+magnetic+resonance+elastography+(microMRE.">Microscopic magnetic resonance elastography (microMRE.</a> Magn. Reson. Med. 54, 605-615 (2005).</li><li>Muthupillai, R., Lomas, D. J., Rossman, P. J., Greenleaf, J. F., Manduca, A., Ehman, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+resonance+elastography+by+direct+visualization+of+propagating+acoustic+strain+waves.">Magnetic resonance elastography by direct visualization of propagating acoustic strain waves.</a> Science. 269, 1854-1857 (1995).</li><li>Othman, S. F., Curtis, E. T., Plautz, S. A., Pannier, A. P., Xu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+resonance+elastography+monitoring+of+tissue+engineered+constructs.">Magnetic resonance elastography monitoring of tissue engineered constructs.</a> NMR Biomed. , (2011).</li><li>Oliphant, T. E., Manduca, A., Ehman, R. L., Greenleaf, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complex-valued+stiffness+reconstruction+for+magnetic+resonance+elastography+by+algebraic+inversion+of+the+differential+equation.">Complex-valued stiffness reconstruction for magnetic resonance elastography by algebraic inversion of the differential equation.</a> Magn. Reson. Med. 45, 299-310 (2001).</li><li>Ringleb, S. I., Chen, Q., Lake, D. S., Manduca, A., Ehman, R. L., An, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+shear+wave:+comparison+to+a+dynamic+shear+material+test.">Quantitative shear wave: comparison to a dynamic shear material test.</a> Magn. Reson. Med. 53, 1197-1201 (2005).</li><li>Hong, L., Peptan, I., Clark, P., Mao, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+vivo+adipose+tissue+engineering+by+human+marrow+stromal+cell+seeded+gelatin+sponge.">Ex vivo adipose tissue engineering by human marrow stromal cell seeded gelatin sponge.</a> Ann. Biomed. Eng. 33, 511-517 (2005).</li><li>Dennis, J. E., Haynesworth, S. E., Young, R. G., Caplan, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteogenesis+in+marrow-derived+mesenchymal+cell+porous+ceramic+composites+transplanted+subcutaneously:+effect+of+fibronectin+and+laminin+on+cell+retention+and+rate+of+osteogenic+expression.">Osteogenesis in marrow-derived mesenchymal cell porous ceramic composites transplanted subcutaneously: effect of fibronectin and laminin on cell retention and rate of osteogenic expression.</a> Cell Transplant. 1, 23-32 (1992).</li><li>Marion, N. W., Mao, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cells+and+tissue+engineering.">Mesenchymal stem cells and tissue engineering.</a> Methods Enzymol. 420, 339-361 (2006).</li><li>Rydberg, J., Grimm, R., Kruse, S., Felmlee, J., McCracken, P., Ehman, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+spin-echo+magnetic+resonance+elastography+of+the+brain.">Fast spin-echo magnetic resonance elastography of the brain.</a> , 1647-1647 (2001).</li><li>Kruse, S. A., Grim, R. C., Lake, D. S., Manduca, A., Ehman, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+EPI+based+3D+MR+elastography+of+the+brain.">Fast EPI based 3D MR elastography of the brain.</a> , 3385-3385 (2006).</li></ol>

I denne fremgangsmåde er fremgangsmåden MRE til manipuleret konstruktioner demonstreret fra cellepræparat til frembringelse af en elastogram. Ved at anvende en ikke-destruktiv mekanisk vurdering af metode til vævsmanipulering rørledningen, er det nu muligt at vurdere ændringer i manipuleret konstruktioner gennem flere stadier af udvikling. Derudover supplerer MRE andre MR-metoder til overvågning manipuleret vævskonstruktioner såsom diffusion, magnetisering overførsel, og kemisk skift analyse 1. ...

<table border="1"><tbody><tr><td> Materialets navn </td><td> Type </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer </td></tr><tr><td> MSCGM-Bullet Kit </td><td> Reagens </td><td> Lonza </td><td> PT-3001 </td><td> Opbevares ved 4 ° C </td></tr><tr><td> 1X DPBS </td><td> Reagens </td><td> Invitrogen </td><td> 21600-010 </td><td></td></tr><tr><td> 0,05% trypsin-EDTA </td><td> Reagens </td><td> Gibco, Invitrogen </td><td> 25300-054 </td><td> Opbevar ved -20 ° C </td></tr><tr><td> Dexamethason </td><td> Reagens </td><td> Sigma-Aldrich </td><td> D2915 </td><td></td></tr><tr><td> 3-isobutyl-1-methylxanthin </td><td> Reagens </td><td> Sigma-Aldrich </td><td> I5879 </td><td> Opbevar ved -20 ° C </td></tr><tr><td> Insulin-bovin pankreas </td><td> Reagens </td><td> Sigma-Aldrich </td><td> I6634 </td><td> Opbevar ved -20 ° C </td></tr><tr><td> Indomethacin </td><td> Reagens </td><td> Sigma-Aldrich </td><td> I7378 </td><td></td></tr><tr><td> Β-glycerophosphat </td><td> Reagens </td><td> Sigma-Aldrich </td><td> G9891 </td><td></td></tr><tr><td> L-ascorbinsyre-2-phosphat </td><td> Reagens </td><td> Sigma-Aldrich </td><td> A8960 </td><td></td></tr><tr><td> Gelfoam </td><td> Scaffold </td><td> Pharmacia &amp; Upjohn Co </td><td> 09-0315-08 </td><td></td></tr><tr><td> Humane mesenchymale stamceller </td><td> Cellelinie </td><td> Lonza </td><td> PT-2501 </td><td></td></tr><tr><td> 9.4T MR Scanner </td><td> Udstyr </td><td> Agilent </td><td></td><td> 400 MHz WB </td></tr><tr><td> 10mm Litz Coil </td><td> Udstyr </td><td> Doty Videnskabelig </td><td></td><td></td></tr><tr><td> Laser Doppler Vibrometer </td><td> Udstyr </td><td> Polytec </td><td> PDV-100 </td><td></td></tr><tr><td> Vibrosoft (20) </td><td> Software </td><td> Polytec </td><td></td><td></td></tr><tr><td> Funktionsgenerator </td><td> Udstyr </td><td> Agilent </td><td> AFG 3022B </td><td></td></tr><tr><td> Forstærker </td><td> Udstyr </td><td> Piezo inc </td><td> EPA-104 til 115 </td><td></td></tr><tr><td> Piezo bukkemotor </td><td> Udstyr </td><td> Piezo inc. </td><td> T234-A4Cl-203X </td><td></td></tr><tr><td> Computer-Linux </td><td> Udstyr </td><td> Processor: Intel Core 2 Duo E8400 Hukommelse: 2G </td><td></td><td></td></tr><tr><td> Computer-Windows </td><td> Udstyr </td><td> Processor: Intel Core 2 Duo E8400 Hukommelse: 2G </td><td></td><td></td></tr><tr><td> MATLAB </td><td> Software </td><td> MathWorks, inc </td><td></td><td> 2009b </td></tr></tbody></table>

magnetic resonance elastography methodology for the evaluation of tissue engineered construct growth

Traditionelle mekaniske afprøvning ofte resulterer i destruktion af prøven, og i tilfælde af længerevarende væv manipuleret konstruktion undersøgelser, er anvendelsen af ​​destruktive vurdering ikke acceptabel. En foreslået alternativ er anvendelsen af ​​en billeddannende proces kaldet magnetisk resonans elastografi. Elastografi er en ikke-destruktiv metode til bestemmelse af konstruerede resultatet ved at måle de lokale mekaniske egenskaber værdier (dvs. komplekse forskydningsmodul), som er vigtige markører til identifikation af struktur og funktionalitet af et væv. Som en noninvasiv midler til evaluering, er overvågningen af manipuleret konstruktioner med billeddiagnostiske metoder såsom magnetisk resonans imaging (MRI) ses en stigende interesse i det seneste årti 1. For eksempel har de magnetisk resonans (MR) teknikker til diffusion og relaxometry været i stand til at karakterisere de ændringer i kemiske og fysiske egenskaber under manipuleret vævsudvikling 2. Den foreslåede metode iFølgende protokol anvendes mikroskopisk magnetisk resonans elastografi (μMRE) som en ikke-invasiv MR baseret teknik til måling af mekaniske egenskaber af små bløde væv 3. MRE opnås ved kobling af en sonisk mekanisk aktuator til vævet af interesse, og registrering af forskydningsbølgen formering med en MR-skanner 4. For nylig har μMRE blevet anvendt i vævsmanipulering at erhverve vigtige vækst, oplysninger, der traditionelt måles ved hjælp af mekaniske destruktive makroskopiske teknikker 5. I det følgende procedure, elastografi opnås ved billeddannelse af gensplejsede konstruktioner med en modificeret Hahn spin-ekko-sekvens koblet med en mekanisk aktuator. Som vist i figur 1, den modificerede sekvens synkroniserer billedoptagelse med transmissionen af ​​eksterne transversalbølger, efterfølgende bevægelse er sensibiliseret ved anvendelse af oscillerende bipolære par. Efter indsamling af billeder med positive og negative bevægelse sensitization, kompleks division af dataene frembringe en forskydningsbølge billede. Derefter bliver billedet vurderes ved hjælp af en inversion algoritme til at generere en forskydningsstivhed kort 6. De resulterende målinger ved hver voxel er blevet vist at korrelere stærkt (r2&gt; 0,9914) med data indsamlet ved hjælp af dynamisk mekanisk analyse 7. I denne undersøgelse er elastografi integreret i vævet udviklingsprocessen for overvågning af humane mesenchymale stamceller (h MSC) differentiering til adipogeniske og osteogene konstruktioner som vist i figur 2.

Watch this Scientific Journal Video about Magnetic Resonance Elastography Methodology for the Evaluation of Tissue Engineered Construct Growth at JoVE.com

Magnetic Resonance Elastography Methodology for the Evaluation of Tissue Engineered Construct Growth

Fremgangsmåden demonstrerer metoden af ​​magnetisk resonans elastografi til overvågning af konstruerede resultatet af fedt-og osteogene manipuleret konstruktioner gennem invasiv lokal vurdering af de mekaniske egenskaber under anvendelse af mikroskopisk magnetisk resonans elastografi (μMRE).

Magnetic Resonance elastografi metode til evaluering af manipuleret væv Construct Vækst

Traditional mechanical testing often results in the destruction of the sample, and in the case of long term tissue engineered construct studies, the use ...

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