Name: in situ compressive loading and correlative noninvasive imaging of the bone-periodontal ligament-tooth fibrous joint
Uploaded: 2014-03-07T13:45:00
Description: Watch this Scientific Journal Video about In situ Compressive Loading and Correlative Noninvasive Imaging of the Bone-periodontal Ligament-tooth Fibrous Joint at JoVE.com

Forfatterne erkender finansieringsstøtte NIH / NIDCR R00DE018212 (SPH), NIH/NIDCR-R01DE022032 (SPH), NIH / NIDCR T32 DE07306 (AJ, JDL), NIH / NCRR S10RR026645, (SPH) og afdelinger for Forebyggende og genoprettende Dental Sciences og orofacial Sciences, UCSF. Desuden Forfatterne erkender Xradia dimittendstipendium (AJ) Xradia Inc., Pleasanton, CA. Forfatterne takker Dr. Kathryn Grandfield, UCSF for hendes hjælp med efterbehandling af data; Drs. Stephen Weiner og Gili Naveh, Weizmann Institute of Science, Rehovot, Israel, Dr. Ron Shahar, The Hebrew University of Jerusalem, Israel for deres indsigtsfulde diskussioner specifikke for in situ læsning enhed. Forfatterne vil også gerne takke biomaterialer og Bioengineering MicroCT Imaging Facility på UCSF for brug af Micro XCT og in situ læsning enhed.

Stalde og eutanasi: Alle dyr, der anvendes i denne demonstration blev huset under patogenfrie betingelser i overensstemmelse med retningslinjerne fra Institutional Animal Care og brug Udvalg (IACUC) og National Institute of Health (NIH). Giv dyrene med standard hard-pille rottefoder og vand ad libitum. Aflive dyr via en to-trins metode til kuldioxid kvælning, bilateral thoracotomi i overensstemmelse med den standard-protokol af UCSF som godkendt af IACUC. Udføre biomekaniske test ...

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Det første skridt i etableringen af ​​denne protokol involveret evaluere stivhed lastrammen ved hjælp af en stiv krop. Baseret på resultaterne, stivhed var signifikant højere muliggør anvendelse af ladeindretningen til yderligere testning af enheder med betydeligt lavere stivhedsværdier. Det andet trin fremhævet evne instrumentet at skelne mellem forskellige stivhedsværdier ved hjælp af to faser af kurven belastning-aflæsning genereres ved brug af et stift legeme, PDMS materialer af forskellige tværbindin...

Adskillige eksperimentelle metoder fortsat anvendes til at undersøge biomekanik diarthrodial og fiber leddene. Specifikke metoder til tand orgel biomekanik omfatter anvendelse af strain gauges 1-3, photoelasticity metoder 4, 5, Moiré interferometri 6, 7, elektronisk pletmønster interferometri 8 og digitalt billede korrelation (DIC) 9-14. I denne undersøgelse, den innovative tilgang omfatter invasiv billeddannelse ved hjælp af røntgenstråler til at afsløre de interne strukturer i et fiberholdigt joint (mineraliseret væv og deres grænseflader består af blødere zoner, og interfacing væv såsom ledbånd) ved belastninger svarende til in vivo. En in situ loading koblet til en mikro-røntgenmikroskop vil blive anvendt. Belastningen tid og load-forskydning kurver vil blive indsamlet som den molære af interesse inden for en frisk høstet rotte hemi-underkæbe er indlæst. Den main mål for den tilgang, der præsenteres i denne undersøgelse er at understrege effekten af ​​tre-dimensionelle morfologi tand-bone ved at sammenligne forholdene på: 1) uden belastning, og når det er indlæst, og når 2) koncentrisk og excentrisk belastet. Eliminerer behovet for afskårne prøver, og til at udføre eksperimenter på hele intakte organer på våd vej vil give mulighed for maksimal bevarelse af 3D stress tilstand. Dette åbner et nyt område for efterforskning i forståelsen dynamiske processer af komplekset under forskellige belastningsforhold scenarier. I denne undersøgelse er de metoder til testning af PDL biomekanik indenfor en intakt fibrøst fælles af en Sprague Dawley rotte, en fælles betragtes som en optimal bioteknik modelsystem vil blive nærmere. Eksperimenter vil omfatte simulering af tygning belastninger under hydreret betingelser for at fremhæve tre vigtige elementer i det fælles, som de vedrører orgel niveau biomekanik. De tre punkter vil omfatte: 1) reaktionær kraft vs forskydning:tand slagvolumen i alveolære sokkel og dens reaktionære reaktion på belastning, 2) tre-dimensionelle (3D) rumlig konfiguration og morphometrics: geometriske forhold af tanden med den alveolære soklen, og 3) ændringer i aflæsning 1 og 2 på grund af en ændring i lastning akse, nemlig fra koncentrisk excentriske belastninger. De tre grundlæggende udlæsninger af den foreslåede teknik kan anvendes til at undersøge den adaptive karakter af fuger i hvirveldyr enten som følge af ændringer i funktionelle krav og / eller sygdom. Ændringer i de førnævnte udlæsninger, specielt sammenhængen mellem reaktionære belastninger med forskydning, og deraf reaktionære load-tids-og load-forskydningskurver ved forskellige Belastningen kan anvendes til at fremhæve de overordnede ændringer i joint biomekanik. Effekten af ​​den foreslåede protokol vil blive evalueret ved at koble mekanisk prøvning udlæsninger til 3D morphometrics og overordnede biomekanik af joint.

<table border="0" cellpadding="0" cellspacing="0" width="652">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Bard Parker Blade</td>
			<td style="width:101px;">BD</td>
			<td style="width:123px;">MEDC-001054</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">AFM metal disk</td>
			<td style="width:101px;">Ted Pella</td>
			<td style="width:123px;">16218</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Polymethyl methacrylate&#160;</td>
			<td style="width:101px;">GC America</td>
			<td style="width:123px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Uni-Etch</td>
			<td style="width:101px;">Bisco</td>
			<td style="width:123px;">E5502EBM</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Optibond Solo Plus</td>
			<td style="width:101px;">Kerr Corp</td>
			<td style="width:123px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Filtek Flow</td>
			<td style="width:101px;">3M</td>
			<td style="width:123px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Hurculite Ultra</td>
			<td style="width:101px;">Kerr</td>
			<td style="width:123px;">34346</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Tris buffer</td>
			<td style="width:101px;">Mediatech Inc.</td>
			<td style="width:123px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Articulating paper</td>
			<td style="width:101px;"></td>
			<td style="width:123px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">Phosphotungstic Acid</td>
			<td style="width:101px;">Sigma Aldrich</td>
			<td style="width:123px;">HT152</td>
			<td></td>
		</tr>
	</tbody>
</table>

in situ compressive loading and correlative noninvasive imaging of the bone-periodontal ligament-tooth fibrous joint

Denne undersøgelse viser en roman biomekanik prøvningsprotokol. Fordelen ved denne protokol indbefatter anvendelsen af en in situ loading koblet til en høj opløsning røntgenmikroskop, hvilket muliggør visualisering af interne strukturelle elementer under simulerede fysiologiske belastninger og våde forhold. Eksperimentelle prøver vil omfatte intakt knogle-parodontal ligament (PDL)-tand fiber leddene. Resultaterne vil illustrere tre vigtige elementer i protokollen, da de kan anvendes til orgel niveau biomekanik: 1) reaktionær kraft vs forskydning: tand slagvolumen i alveolære sokkel og dens reaktionære reaktion på belastning, 2) tre-dimensionelle (3D) rumlige konfiguration og morphometrics: geometriske forhold af tanden med den alveolære soklen, og 3) ændringer i aflæsning 1 og 2 på grund af en ændring i lastning akse, nemlig fra koncentrisk excentriske laster. Effekten af ​​den foreslåede protokol vil blive evalueret ved at koble mekanisk testing udlæsninger til 3D morphometrics og overordnede biomekanik af samlingen. Desuden vil denne teknik understrege behovet for at komme i ligevægt eksperimentelle betingelser, specielt reaktionære belastninger forud for erhvervelse tomogrammer fiberholdigt leddene. Det skal bemærkes, at den foreslåede protokol er begrænset til afprøvning præparater under ex vivo-betingelser, og at anvendelse af kontrastmidler til at visualisere blødt væv mekanisk reaktion kan føre til fejlagtige konklusioner om vævs-og organ-niveau biomekanik.

Estimering af lastning enhed &quot;backlash&quot;, &quot;pushback&quot;, stivhed og systemet afdrift under en konstant belastning Backlash: Mellem lastning og losning dele af cyklen, findes der en pause på 3 sek hvorunder gear omvendt i motoren før sand aflæsning påbegyndes, nemlig som prøven trækker sig væk fra den øverste kæbe (Figur 3). Denne periode kaldes et tilbageslag i systemet, som udgør et segment af tid, når sy...

Watch this Scientific Journal Video about In situ Compressive Loading and Correlative Noninvasive Imaging of the Bone-periodontal Ligament-tooth Fibrous Joint at JoVE.com

In situ Compressive Loading and Correlative Noninvasive Imaging of the Bone-periodontal Ligament-tooth Fibrous Joint

I denne undersøgelse, vil brugen af en in situ loading koblet med mikro-X-ray computertomografi for fiber joint biomekanik blive diskuteret. Eksperimentelle udlæsninger identificerbare med en generel ændring i fælles biomekanik vil omfatte: 1) reaktionær kraft vs forskydning, dvs tand slagvolumen i alveolære sokkel og dens reaktionære reaktion på belastning, 2) tre-dimensionelle (3D) rumlig konfiguration og morphometrics, dvs geometriske Forholdet af tanden med den alveolære soklen, og 3) ændringer i udlæsninger 1 og 2 på grund af en ændring i lastning akse, dvs, koncentriske eller excentriske belastninger.

In situ trykbelastning og korrelationsmaalinger Noninvasiv Imaging af Bone-parodontal ligament-tand Fiber Joint

This study demonstrates a novel biomechanics testing protocol. The advantage of this protocol includes the use of an in situ loading device coupled to a ...

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Bioengineering

In situ Compressive Loading and Correlative Noninvasive Imaging of the Bone-periodontal Ligament-tooth Fibrous Joint

High-resolution Fiber-optic Microendoscopy for in situ Cellular Imaging

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Undersøgelse af støkiometrien af ​​epidermal vækstfaktor receptor i intakte celler ved hjælp af korrelative Microscopy

This protocol describes the labeling of epidermal growth factor receptor (EGFR) on COS7 fibroblast cells, and subsequent correlative fluorescence ...

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles

Korrelative lys- og Electron Microscopy Brug Quantum Dot Nanopartikler

A method is described whereby quantum dot (QD) nanoparticles can be used for correlative immunocytochemical studies of human pathology tissue using ...

Characterizing Cell Migration Within Three-dimensional In Vitro Wound Environments

Characterizing Cell Migration Within Three-dimensional In Vitro Wound Environments

Kendetegner celle Migration inden for tre-dimensionelle In Vitro sår miljøer

Currently, most in vitro models of wound healing, such as well-established scratch assays, involve studying cell migration and wound closure on ...

Optimized Setup and Protocol for Magnetic Domain Imaging with In Situ Hysteresis Measurement

Optimized Setup and Protocol for Magnetic Domain Imaging with In Situ Hysteresis Measurement

Optimeret installation og protokol for magnetisk domæne Imaging med In Situ hysterese måling

This paper elaborates the sample preparation protocols required to obtain optimal domain patterns using the Bitter method, focusing on the extra steps ...

Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics

Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics

Sensorer til at registrere Protease aktivitet In Vivo for invasiv diagnostik

Proteases are multi-functional enzymes that specialize in the hydrolysis of peptide-bonds and control broad biological processes including homeostasis and ...