Name: proximal cadaveric femur preparation for fracture strength testing and quantitative ct-based finite element analysis
Uploaded: 2017-03-11T13:00:00
Description: Watch this Scientific Journal Video about Proximal Cadaveric Femur Preparation for Fracture Strength Testing and Quantitative CT-based Finite Element Analysis at JoVE.com

Vi vil gjerne takke Materialer og strukturell testing Kjerne Facility ved Mayo Clinic for teknisk support. I tillegg ønsker vi å takke Lawrence J. Berglund, Brant Newman, Jorn op den Buijs, Ph.D., for deres hjelp i løpet av studien. Denne studien ble finansielt støttet av Grainger Innovation Fund fra Grainger Foundation.

MERK: Alle studier presentert i denne protokollen ble godkjent av Institutional Review Board (IRB) ved Mayo Clinic. Benene ble oppnådd over en periode på 6 år fra ulike organisasjoner. Alle prøvene ble samlet inn i løpet av 72 timer etter døden, innpakket i saltvann mettet håndklær, og lagret ved -20 ° C til forberedelse. 1. Måling bentetthet ved bruk av DXA <ol><li> Fjerne prøvene holdt ved -20 ° C fryser for å tine ved romtemperatur i ca. 24 timer; prøvestykker behøver ikk...

<ol>
	<li>Cristofolini, L., Schileo, E., Juszczyk, M., Taddei, F., Martelli, S., Viceconti, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+testing+of+bones:+the+positive+synergy+of+finite-element+models+and+in+vitro+experiments.">Mechanical testing of bones: the positive synergy of finite-element models and in vitro experiments.</a> Philos Trans A Math Phys Eng Sci. 368, 2725-2763 (2010).</li><li>Cartner, J. L., Hartsell, Z. M., Ricci, W. M., Tornetta, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+we+trust+ex+vivo+mechanical+testing+of+fresh-frozen+cadaveric+specimens?+The+effect+of+postfreezing+delays.">Can we trust ex vivo mechanical testing of fresh-frozen cadaveric specimens? The effect of postfreezing delays.</a> J Orthop Trauma. 25 (8), 459-461 (2011).</li><li>An, Y. H., Draughn, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mechanical testing of bone and the bone-implant interface. , (1999).</li><li>van Haaren, E. H., van der Zwaard, B. C., van der Veen, A. J., Heyligers, I. C., Wuisman, P. I., Smit, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+long-term+preservation+on+the+mechanical+properties+of+cortical+bone+in+goats.">Effect of long-term preservation on the mechanical properties of cortical bone in goats.</a> Acta Orthop. 79, 708-716 (2008).</li><li>Shaw, J. M., Hunter, S. A., Gayton, J. C., Boivin, G. P., Prayson, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeated+freeze-thaw+cycles+do+not+alter+the+biomechanical+properties+of+fibular+allograft+bone.">Repeated freeze-thaw cycles do not alter the biomechanical properties of fibular allograft bone.</a> Clin Orthop Relat Res. 470 (3), 937-943 (2012).</li><li>Topp, T., 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=Embalmed+and+fresh+frozen+human+bones+in+orthopedic+cadaveric+studies:+which+bone+is+authentic+and+feasible?.">Embalmed and fresh frozen human bones in orthopedic cadaveric studies: which bone is authentic and feasible?.</a> Acta Orthop. 83 (5), 543-547 (2012).</li><li>Manske, 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=Cortical+and+trabecular+bone+in+the+femoral+neck+both+contribute+to+proximal+femur+failure+load+prediction.">Cortical and trabecular bone in the femoral neck both contribute to proximal femur failure load prediction.</a> Osteoporos Int. 20 (3), 445-453 (2009).</li><li>Rezaei, A., Dragomir-Daescu, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femoral+Strength+Changes+Faster+With+Age+Than+BMD+in+Both+Women+and+Men:+A+Biomechanical+Study.">Femoral Strength Changes Faster With Age Than BMD in Both Women and Men: A Biomechanical Study.</a> J Bone Miner Res. 30, 2200-2206 (2015).</li><li>Cristofolini, L., McNamara, B., Freddi, A., Viceconti, M. <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+measured+strains+in+the+loaded+femur:+quantification+of+experimental+error.">In vitro measured strains in the loaded femur: quantification of experimental error.</a> J Strain Anal Eng Des. 32, 193-200 (1997).</li><li>Dragomir-Daescu, D., 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=Robust+QCT/FEA+models+of+proximal+femur+stiffness+and+fracture+load+during+a+sideways+fall+on+the+hip.">Robust QCT/FEA models of proximal femur stiffness and fracture load during a sideways fall on the hip.</a> Ann Biomed Eng. 39, 742-755 (2011).</li></ol>

Vi presenterte en robust bein forberedelse protokoll for å sikre mekanisk testing og QCT / FEA modellering av femoral styrke i en sidelengs fall på hoften konfigurasjon. Denne metoden ble vår standard in-house-protokollen. I løpet av 6 år, med varierende personell, ca 200 lårben ble vellykket fremstilt etter denne protokollen. Resultatene av protokollen omfatter klassifisere bein forhold ved hjelp av DXA, utelukker metastatisk sykdom, tidligere frakturer, eller implantater ved hjelp av røntgen, og skaffe mineral ...

Determining the true cadaveric proximal femoral fracture strength with mechanical testing is a destructive method that requires a rigorous testing approach for accurate measurements. In particular, proper bone preparation methods are necessary to maintain near in vivo integrity of the bones prior to mechanical fracture testing1. This is achieved by proper bone storage and minimizing handling at room temperature. This test data is extensively used to validate QCT/FEA models of femoral fracture which have the potential to be used clinically to understand the fracture risk, especially in osteoporotic patients. Unfortunately, there is no current standard procedure to prepare proximal femur samples for mechanical testing. A good testing procedure should ensure repeatability and reproducibility of the preparation process. Therefore, fixtures required for sample preparation need to be carefully designed and fabricated to minimize the likelihood of various testing errors. We also need to minimize the preparation time for which bone tissue is at room temperature and thus in danger of degradation with irreversible changes in mechanical properties. 
To this end, we have developed a procedure that preserves bone tissue across multiple preparation steps. This is important to ensure minimal exposure time at room temperature while also minimizing the number of freeze/thaw cycles which can affect tissue physical properties2. The entire procedure is long and nontrivial as the steps occurred over multiple weeks and required scheduling for scanning procedures and personnel availability. The steps included thawing bone samples, screening the samples using DXA scanning to obtain bone mineral density (BMD) values, X-ray to rule out any diseased specimens, and finally CT scanning to estimate distribution of bone mineral and femoral geometry. All the specimens were prepared for testing by removing extraneous soft tissues from the bone surface, cutting the femur to a length required for testing, and potting the femur in a desired orientation for simulating a sideways fall on the hip during subsequent testing. It is essential to keep the time period for all these operations as short as possible. A robust protocol is thus mandatory for consistent specimen preparation, tissue preservation between steps, and for reducing the overall preparation time. 
The aim of this paper is to present in detail the procedures involved in the preparation of femoral samples for subsequent mechanical testing under various conditions. Preservation of the bone tissue is crucial in this process and we achieved it by keeping specimens frozen between steps and keeping them carefully wrapped in saline saturated towels at all times except when scanning and mechanically testing the bones. Femora were also kept wrapped in saline wet towels during the steps involving PMMA curing to prevent dryness of the bone tissue.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CT potting container and scanning fixture</td>
			<td>Internally manufactured</td>
			<td>N/A</td>
			<td>Custom designed and manufactured</td>
		</tr>
		<tr>
			<td>CT scanner</td>
			<td>Siemens</td>
			<td>Somatom Definition scanner (Siemens, Malvern, PA)</td>
			<td>CT scanning equipment</td>
		</tr>
		<tr>
			<td>Quantitative CT Phantom</td>
			<td>Midways Inc, San Francisco, CA</td>
			<td>Model 3 CT calibration Phantom</td>
			<td>Used for obtaining BMD values from Hounsfield units in the CT image</td>
		</tr>
		<tr>
			<td>Dual Energy X-ray Absorptiometry scanner</td>
			<td>General Electric</td>
			<td>N/A</td>
			<td>GE Lunar iDXA scanner for bone health or any similar BMD scanners</td>
		</tr>
		<tr>
			<td>Hygenic Orhodontic Resin (PMMA)</td>
			<td>Patterson Dental Supply</td>
			<td>H02252</td>
			<td>Controlled substance and can be purchased with proper approval</td>
		</tr>
		<tr>
			<td>Freezer</td>
			<td>Kenmore</td>
			<td>N/A</td>
			<td>This is a -20oC storage for bones</td>
		</tr>
		<tr>
			<td>X-ray scanner</td>
			<td>General Electric</td>
			<td>&#160;46-270615P1</td>
			<td>X-ray imaging equipment.&#160;&#160;</td>
		</tr>
		<tr>
			<td>X-ray films</td>
			<td>Kodak</td>
			<td>N/A</td>
			<td>Used to display x-ray images</td>
		</tr>
		<tr>
			<td>X-ray developer</td>
			<td>Kodak X-Omatic</td>
			<td>M35A X-OMAT&#160;</td>
			<td>Used for developing X-ray images</td>
		</tr>
		<tr>
			<td>X-ray Cassette</td>
			<td>Kodak X-Omatic</td>
			<td>N/A</td>
			<td>Used for holding x-ray films</td>
		</tr>
		<tr>
			<td>5-pound Rice Bags</td>
			<td>Great Value</td>
			<td>N/A&#160;</td>
			<td>Used for mimicking soft tissue during the DXA scanning process</td>
		</tr>
		<tr>
			<td>Physiologic Saline (0.9% Sodium Chloride)</td>
			<td>Baxter</td>
			<td>NDC 0338-0048-04</td>
			<td>Used for keeping samples hydrated</td>
		</tr>
		<tr>
			<td>Scalpels and scrapers</td>
			<td>Bard-Parker</td>
			<td>N/A</td>
			<td>Used to clean the bone from soft tissue</td>
		</tr>
		<tr>
			<td>Cast cutter</td>
			<td>Stryker</td>
			<td>810-BD001</td>
			<td>Used to cut femoral shaft</td>
		</tr>
		<tr>
			<td>Drilling machine</td>
			<td>Bosch</td>
			<td>N/A</td>
			<td>Used to drill the femoral shaft</td>
		</tr>
		<tr>
			<td>Fume Hood</td>
			<td>Hamilton</td>
			<td>70532</td>
			<td>Used for ventilation when using making PMMA</td>
		</tr>
	</tbody>
</table>

proximal cadaveric femur preparation for fracture strength testing and quantitative ct-based finite element analysis

Avdød brudd testing er rutinemessig brukt til å forstå hvilke faktorer som påvirker proksimale femur styrke. Fordi ex vivo biologisk vev er tilbøyelige til å miste sine mekaniske egenskaper over tid, prøven forberedelse for eksperimentell testing må utføres nøye for å få pålitelige resultater som representerer in vivo forhold. Av den grunn har vi utviklet en protokoll og et sett av oversikten for å fremstille de femorale prøvene slik at deres mekaniske egenskaper opplevd minimale endringer. De lårben ble holdt i en frossen tilstand bortsett fra under forberedelse trinn og mekanisk testing. De relevante kliniske tiltak av total hofte og lårhals beinmineraltetthet (BMD) ble oppnådd med en klinisk dual X-ray absorpsjonsmetri (DXA) bein densitometer, og 3D-geometri og distribusjon av beinmineral ble oppnådd ved bruk av CT med en kalibrerings fantom for kvantitative beregninger basert på gråtone verdier. Enhver mulig bensykdom, brudd, Eller tilstedeværelse av implantater eller gjenstander som påvirker beinstrukturen, ble utelukket med røntgenskanning. For forberedelse, ble alle bein nøye renset av overflødig bløtvevet, og ble kuttet og puttet på intern rotasjon vinkel av interesse. Et skjære lampen tillates den distale enden av benet som skal kappes av forlater den proksimale femur ved en ønsket lengde. Å tillate posisjonering av lårhalsen på foreskrevet vinkler under senere CT scanning og mekanisk testing, ble de proksimale femoral skaft puttet i polymethylmethacrylate (PMMA) med en armatur utviklet spesielt for ønskede orienteringer. De oppsamlede fra våre eksperimenter data ble deretter brukt for validering av kvantitativ computer tomografi (QCT) -baserte finite element analyse (FEA), som beskrevet i en annen protokoll. I dette manuskriptet, presenterer vi protokollen for nøyaktig bein forberedelse for mekanisk testing og påfølgende QCT / FEA modellering. Den nåværende protokollen ble vellykket anvendt for å forberede ca 200 cadaveric lårben over en 6-års periode.

De avdød lårben ble sendt frosset og holdt ved -20 ° C inntil fremstillingen begynte. BMD skanning ble utført ved hjelp av en DXA skanner for å måle total hofte og nakke BMD samt T-score for hver prøve (figur 1). En T-score er antall standardavvik av den målte BMD sammenlignet med gjennomsnittsverdier for unge, friske personer. Det kan være alt fra -2,5 eller lavere for osteoporotiske ben, mellom -1 og -2,5 til osteopene ben og høyere enn for vanlige -1 bein. N...

Watch this Scientific Journal Video about Proximal Cadaveric Femur Preparation for Fracture Strength Testing and Quantitative CT-based Finite Element Analysis at JoVE.com

Proximal Cadaveric Femur Preparation for Fracture Strength Testing and Quantitative CT-based Finite Element Analysis

Vi presenterer en robust protokoll om hvordan å nøye bevare og forberede avdød lårben for brudd testing og kvantitativ computertomografi bildebehandling. Metoden gir presis kontroll over inngangsvilkår for det formål å bestemme forholdet mellom bentetthet, bruddstyrke, og definere finite element modellen geometri og egenskaper.

Proksimale avdød Femur Forberedelse til Fracture Styrke Testing og kvantitativ CT-basert Finite Element Analysis

Cadaveric fracture testing is routinely used to understand factors that affect proximal femur strength.  Because ex vivo biological tissues are prone to ...

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