Name: Forfatter Spotlight: Integration af mekanisk og biologisk analyse i tendinopatiforskning
Uploaded: 2024-03-01T13:00:00
Description: Watch this Scientific Journal Video about Integrating Mechanical and Biological Analysis in Tendinopathy Research at JoVE.com

Vi vil gerne anerkende vores finansieringsstøtte: Joe Fallon Research Fund, Dr. Louis Meeks BIDMC Sports Medicine Trainee Research Fund og et intramuralt tilskud (AN), alle fra BIDMC Ortopædi, sammen med støtte fra National Institutes of Health (2T32AR055885 (PMW)).

Denne undersøgelse blev udført pr. Institutional Animal Care and Use Committee (IACUC) godkendelse på Beth Israel Deaconess Medical Center. Dyrene blev bedøvet med 5% isofluran til induktion og 2,5% til vedligeholdelse, og man sørgede for at undgå hypotermi.
1. Opsætning af testsystemet
<ol><li>Kontroller passiv ankelrotation med en stepmotor for at anvende ensartet rotation og drejningsmoment. Styr stepmotoren med en mikrocontroller. Brug input fra 3D-positionen og o...

Cyklisk mekanisk belastning til fremkaldelse af tendinopati i rotteakillessene

<ol>
	<li>Kaux, J. F., Forthomme, B., Goff, C. L., Crielaard, J. M., Croisier, 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=Current+opinions+on+tendinopathy.">Current opinions on tendinopathy.</a> J Sports Sci Med. 10 (2), 238-253 (2011).</li><li>Maffulli, N., Longo, U. G., Kadakia, A., Spiezia, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Achilles+tendinopathy.">Achilles tendinopathy.</a> Foot Ankle Surg. 26 (3), 240-249 (2020).</li><li>Teunis, T., Lubberts, B., Reilly, B. T., Ring, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+and+pooled+analysis+of+the+prevalence+of+rotator+cuff+disease+with+increasing+age.">A systematic review and pooled analysis of the prevalence of rotator cuff disease with increasing age.</a> J Shoulder Elbow Surg. 23 (12), 1913-1921 (2014).</li><li>von Rickenbach, K. J., Borgstrom, H., Tenforde, A., Borg-Stein, J., McInnis, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Achilles+tendinopathy:+evaluation,+rehabilitation,+and+prevention.">Achilles tendinopathy: evaluation, rehabilitation, and prevention.</a> Curr Sports Med Rep. 20 (6), 327-334 (2021).</li><li>Aicale, R., Oliviero, A., Maffulli, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+Achilles+and+patellar+tendinopathy:+what+we+know,+what+we+can+do.">Management of Achilles and patellar tendinopathy: what we know, what we can do.</a> J Foot Ankle Res. 13 (1), 59 (2020).</li><li>Jarvinen, T. A., 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=Achilles+tendon+injuries.">Achilles tendon injuries.</a> Curr Opin Rheumatol. 13 (2), 150-155 (2001).</li><li>Silbernagel, K. G., Hanlon, S., Sprague, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+clinical+concepts:+conservative+management+of+Achilles+tendinopathy.">Current clinical concepts: conservative management of Achilles tendinopathy.</a> J Athl Train. 55 (5), 438-447 (2020).</li><li>Tayfur, A., 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=Are+landing+patterns+in+jumping+athletes+associated+with+patellar+tendinopathy?+A+systematic+review+with+evidence+gap+map+and+meta-analysis.">Are landing patterns in jumping athletes associated with patellar tendinopathy? A systematic review with evidence gap map and meta-analysis.</a> Sports Med. 52 (1), 123-137 (2022).</li><li>Malliaras, P., Cook, J. L., Kent, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+ankle+dorsiflexion+range+may+increase+the+risk+of+patellar+tendon+injury+among+volleyball+players.">Reduced ankle dorsiflexion range may increase the risk of patellar tendon injury among volleyball players.</a> J Sci Med Sport. 9 (4), 304-309 (2006).</li><li>Backman, L. J., Danielson, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+range+of+ankle+dorsiflexion+predisposes+for+patellar+tendinopathy+in+junior+elite+basketball+players:+a+1-year+prospective+study.">Low range of ankle dorsiflexion predisposes for patellar tendinopathy in junior elite basketball players: a 1-year prospective study.</a> Am J Sports Med. 39 (12), 2626-2633 (2011).</li><li>Glazebrook, M. A., Wright, J. R., Langman, M., Stanish, W. D., Lee, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+analysis+of+achilles+tendons+in+an+overuse+rat+model.">Histological analysis of achilles tendons in an overuse rat model.</a> J Orthop Res. 26 (6), 840-846 (2008).</li><li>Carpenter, J. E., Flanagan, C. L., Thomopoulos, S., Yian, E. H., Soslowsky, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+overuse+combined+with+intrinsic+or+extrinsic+alterations+in+an+animal+model+of+rotator+cuff+tendinosis.">The effects of overuse combined with intrinsic or extrinsic alterations in an animal model of rotator cuff tendinosis.</a> Am J Sports Med. 26 (6), 801-807 (1998).</li><li>Gao, H. G., 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=Increased+serum+and+musculotendinous+fibrogenic+proteins+following+persistent+low-grade+inflammation+in+a+rat+model+of+long-term+upper+extremity+overuse.">Increased serum and musculotendinous fibrogenic proteins following persistent low-grade inflammation in a rat model of long-term upper extremity overuse.</a> PLoS One. 8 (8), e71875 (2013).</li><li>Fung, 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=Early+response+to+tendon+fatigue+damage+accumulation+in+a+novel+in+vivo+model.">Early response to tendon fatigue damage accumulation in a novel in vivo model.</a> J Biomech. 43 (2), 274-279 (2010).</li><li>Ueda, Y., 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=Molecular+changes+to+tendons+after+collagenase-induced+acute+tendon+injury+in+a+senescence-accelerated+mouse+model.">Molecular changes to tendons after collagenase-induced acute tendon injury in a senescence-accelerated mouse model.</a> BMC Musculoskelet Disord. 20 (1), 120 (2019).</li><li>Bloom, E., 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=Overload+in+a+rat+in+vivo+model+of+synergist+ablation+induces+tendon+multi-scale+structural+and+functional+degeneration.">Overload in a rat in vivo model of synergist ablation induces tendon multi-scale structural and functional degeneration.</a> J Biomech Eng. 145 (8), 081003 (2023).</li><li>Williamson, P. 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+passive+ankle+dorsiflexion+testing+system+to+assess+mechanobiological+and+structural+response+to+cyclic+loading+in+rat+Achilles+tendon.">A passive ankle dorsiflexion testing system to assess mechanobiological and structural response to cyclic loading in rat Achilles tendon.</a> J Biomech. 156, 111664 (2023).</li><li>Oliveira, L. F., Peixinho, C. C., Silva, G. A., Menegaldo, L. 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+vivo+passive+mechanical+properties+estimation+of+Achilles+tendon+using+ultrasound.">In vivo passive mechanical properties estimation of Achilles tendon using ultrasound.</a> J Biomech. 49 (4), 507-513 (2016).</li></ol>

Denne undersøgelse præsenterer en metode til cyklisk belastning af rottens akillessene med et passivt ankeldorsiflexionsystem til en in vivo overforbrugsinduceret tendinopati-model. Systemets betydning ligger i dets evne til at isolere akillessenen, anvende kvantificerbare belastninger uden kirurgisk adgang til senen og måle in vivo seneegenskaber. 
I 2010 præsenterede Fung et al. en rottepatellar senetræthedsmodel med et specialbygget testsystem14</sup...

Da sener forbinder muskler til knogler og oplever daglige gentagne bevægelser gennem hele deres levetid, er de meget tilbøjelige til overbelastningsskader, der er smertefulde og begrænsende og resulterer i nedsat mekanisk funktion, der påvirker 30-50% af befolkningen1. Tendinopatier er kroniske tilstande, der betragtes som overbelastningsskader på grund af gentagne træthedsbevægelser og utilstrækkelig heling til niveauet før skaden. Både øvre og nedre ekstremiteter er almindeligt påvirket, herunder rotator manchet, albue, akillessene, og patellar senen 2,3,4,5. Akillessenopati er almindelig i aktiviteter, der involverer løb og spring, især atleter involveret i atletik, mellem- og langdistanceløb, tennis og andre boldsport, der påvirker 7-9% af løberne 6,7. Skader fra løb og spring kan også forårsage begrænset ankeldorsiflexion, hvilket er en risikofaktor for Achilles og patellar tendinopathies 8,9,10. Der er således behov for en bedre vurdering og karakterisering af tendinopati, som denne undersøgelse kan give som en rottemodel af passiv ankeldorsiflexion til overforbrug af akillesseneskader.
Tidligere arbejde med små dyremodeller har været rettet mod at studere udviklingen og markørerne for tendinopati. Disse omfatter løbebånd øvelse, gentagne rækkevidde, direkte sene belastning, kollagenaseinjektioner, kirurgi, og in vitro undersøgelser 11,12,13,14,15,16. Selvom litteraturen har draget fordel af identifikationen af skademarkører ved at anvende disse tendinopatimodeller, omfatter begrænsninger belastning af senen i ikke-fysiologisk relevante ledbevægelser, som i tilfælde af direkte belastning af senen, ikke direkte måling af påførte belastninger, såsom til løbebåndsundersøgelser, og ikke anvendelse af fysiologisk overforbrug, som i tilfældet med kollagenaseinjektioner, blandt andre. Til dette formål havde denne undersøgelse til formål at udvikle et system, der noninvasivt anvender kvantificerede belastninger på akillessenen med anvendelse af overforbrugsinducerede tendinopatiundersøgelser for at udfylde hullerne i tidligere udviklede smådyrsmodeller for tendinopati. Vi udførte en pilotundersøgelse for at demonstrere, at systemet inducerer reproducerbare ændringer i mekaniske egenskaber over en række belastningscyklusser. Dette system gør det muligt for fysiologisk relevant bevægelse og belastning at fremkalde overforbrug, samtidig med at det kvantificerer og måler de kræfter, der påføres og opleves af senen under belastningsregimet.

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			<td>1/32&#39;&#39; Aluminum beads</td>
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			<td>2.5% isoflurane</td>
			<td></td>
			<td></td>
			<td></td>
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			<td>3D digitizing pen</td>
			<td>Polhemus, Vermont, NH, USA</td>
			<td></td>
			<td></td>
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			<td>3D electromagnetic positioning and orientation sensor</td>
			<td>Polhemus, Vermont, NH, USA</td>
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			<td>5% isoflurane</td>
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			<td>Customized device: 1) Assembly, sensors, 3D printed animal bed and ankle mount actuator</td>
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			<td>Assembled as described in manuscript</td>
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			<td>MATLAB code</td>
			<td>MATLAB, Natick, MA, USA</td>
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			<td>Microcontroller</td>
			<td>Ivrea, Italy</td>
			<td>Arduino UNO, Rev3&#160;</td>
			<td></td>
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			<td>Nose cone</td>
			<td></td>
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			<td>Scalpel and scalpel holder</td>
			<td></td>
			<td></td>
			<td>No. 11 scalpel</td>
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			<td>Sprague-Dawley rats</td>
			<td>Charles River Laboratories, Wilmington, MA, USA</td>
			<td></td>
			<td>11-13 weeks old</td>
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			<td>Stepper driver</td>
			<td>SparkFun Electronics, Niwot, CO 80503</td>
			<td>DM542T</td>
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			<td>Stepper motor</td>
			<td>SparkFun Electronics, Niwot, CO 80503</td>
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			<td>Torque sensor assembly</td>
			<td>Futek Inc., Irvine, CA, USA&#160;</td>
			<td>FSH03985, FSH04473, FSH03927</td>
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			<td>Water heating pad</td>
			<td></td>
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a passive ankle dorsiflexion testing system for an in vivo model of overuse-induced tendinopathy

Tendinopati er en kronisk senetilstand, der resulterer i smerter og tab af funktion og skyldes gentagen overbelastning af senen og begrænset restitutionstid. Denne protokol beskriver et testsystem, der cyklisk påfører mekaniske belastninger via passiv dorsiflexion på rottens akillessene. Den specialskrevne kode består af præ- og postcykliske belastningsmålinger for at vurdere virkningerne af belastningsprotokollen sammen med feedback-kontrolbaseret cyklisk træthedsbelastningsregime. 
Vi brugte 25 Sprague-Dawley-rotter til denne undersøgelse, hvor 5 rotter pr. Gruppe modtog enten 500, 1.000, 2.000, 3.600 eller 7.200 cyklusser med træthedsbelastninger. De procentvise forskelle mellem de præ- og postcykliske belastningsmålinger af hysteresen, spidsbelastningen og belastnings- og losningsmodulerne blev beregnet. Resultaterne viser, at systemet kan forårsage varierende grader af skade på akillessenen baseret på antallet af påførte belastninger. Dette system tilbyder en innovativ tilgang til at anvende kvantificerede og fysiologisk varierende grader af cykliske belastninger på akillessenen til en in vivo-model af træthedsinduceret overforbrugsseneskade.

As tendons connect muscle to bone and experience daily repetitive motions throughout their lifetime, they are highly prone to overuse injuries that are painful and limiting and result in impaired mechanical function, affecting 30-50% of the population1. Tendinopathies are chronic conditions considered overuse injuries due to repetitive fatigue motions and inadequate healing to pre-injury levels. Both upper and lower extremities are commonly affected, including the rotator cuff, elbow, Achilles tendon, and patellar tendon2,3,4,5. Achilles tendinopathy is common in activities involving running and jumping, especially athletes involved in track and field, middle- and long-distance running, tennis, and other ball sports, affecting 7-9% of runners6,7. Injuries from running and jumping may also cause limited ankle dorsiflexion, which is a risk factor for Achilles and patellar tendinopathies8,9,10. Thus, there is a need for a better assessment and characterization of tendinopathy, which this study can provide as a rat model of passive ankle dorsiflexion for overuse Achilles tendon injuries.
Previous work using small animal models has been aimed at studying the development and markers of tendinopathy. These include treadmill exercise, repetitive reaching, direct tendon loading, collagenase injections, surgery, and in vitro studies11,12,13,14,15,16. Although the literature has benefited from the identification of damage markers from employing these tendinopathy models, limitations include loading the tendon in non-physiologically relevant joint motions, as in the case of direct loading of the tendon, not directly measuring applied loads, such as for treadmill studies, and not using physiological overuse, as in the case for collagenase injections, among others. To that end, this study aimed to develop a system that noninvasively applies quantified loads to the Achilles tendon with the application for overuse-induced tendinopathy studies to fill the gaps in previously developed small animal models for tendinopathy. We performed a pilot study to demonstrate that the system induces reproducible changes in mechanical properties over a range of loading cycles. This system enables physiologically relevant motion and loading to induce overuse while simultaneously quantifying and measuring the forces applied to and experienced by the tendon during the loading regimen.

Forfatter Spotlight: Integration af mekanisk og biologisk analyse i tendinopatiforskning 

Et passivt ankeldorsiflexion-testsystem til en in vivo-model af overforbrugsinduceret tendinopati

Our work addresses the mechano biological factors involved in the initiation and progression of overuse induced tendinopathy. The questions we are trying to answer include what factors of exercise, such as frequency, magnitude, and duration of load affect the tendon's response to injury. Current tools such as mechanical measures, histology, and biological assays to evaluate outcome measures are centered and limited to end stage tissue harvesting.With this model, we can explore in vivo continuous measurements with tools such as ultrasound and MRI In the field, in vivo models load the tendon and physiologically relevant manners, while ex vivo models allow for the measurement of biological responses to injuries. Our protocol combines these two measures for direct measurement of tendon stresses and strains during injuries and the associated biological changes. We've established an adaptive system to induce tendon injuries, which allows for simultaneous exploration of mechanical and biological factors.Having established this model, future directions include assessing different parameters, therapeutic avenues, and rehabilitation measures.

Med det stigende antal anvendte cyklusser var der en større reduktion i in vivo senemekaniske egenskaber. Der var en signifikant lavere reduktion i hysterese og laste- og losningsmoduli for 500-cyklusgruppen sammenlignet med 3.600- og 7.200-cyklusgrupperne (p < 0,05) (figur 2). Mens der var en signifikant reduktion i peak stress pr. cyklus fra 500 cyklus til 3.600 cyklus gruppen, var der ingen signifikant reduktion mellem 500 og 7.200 cyklus grupperne. Der var et konsekvent procent...

Watch this Scientific Journal Video about Integrating Mechanical and Biological Analysis in Tendinopathy Research at JoVE.com

Denne protokol præsenterer et testsystem, der anvendes til at fremkalde kvantificerbare og kontrollerede træthedsskader i en rotteakillessene for en in vivo-model af overforbrugsinduceret tendinopati. Proceduren består i at fastgøre rottens ankel til en fælles aktuator, der udfører passiv ankeldorsiflexion med et specialskrevet MATLAB-script.

Tendinopathy is a chronic tendon condition that results in pain and loss of function and is caused by repeated overload of the tendon and limited recovery ...

Vores arbejde omhandler de mekaniske biologiske faktorer, der er involveret i initiering og progression af overforbrugsinduceret tendinopati. De spørgsmål, vi forsøger at besvare, omfatter, hvilke træningsfaktorer, såsom hyppighed, størrelse og varighed af belastning, der påvirker senens reaktion på skade. Nuværende værktøjer såsom mekaniske foranstaltninger, histologi og biologiske assays til evaluering af resultatmål er centreret og begrænset til vævshøstning i slutstadiet.Med denne model kan vi udforske in vivo kontinuerlige målinger med værktøjer som ultralyd og MR I marken belaster in vivo-modeller senen og fysiologisk relevante manerer, mens ex vivo-modeller giver mulighed for måling af biologiske reaktioner på skader. Vores protokol kombinerer disse to mål til direkte måling af senespændinger og belastninger under skader og de tilhørende biologiske ændringer. Vi har etableret et adaptivt system til at fremkalde seneskader, som giver mulighed for samtidig udforskning af mekaniske og biologiske faktorer.Efter at have etableret denne model omfatter fremtidige retninger vurdering af forskellige parametre, terapeutiske veje og rehabiliteringsforanstaltninger.

a-passive-ankle-dorsiflexion-testing-system-for-an-vivo-model-overuse

Research

JoVE Journal

Bioengineering

A Passive Ankle Dorsiflexion Testing System for an In Vivo Model of Overuse-induced Tendinopathy

Tendinopathy is a chronic tendon condition that results in pain and loss of function and is caused by repeated overload of the tendon and limited recovery time. This protocol describes a testing system that cyclically applies mechanical loads via passive dorsiflexion to the rat Achilles tendon. The custom-written code consists of pre- and post-cyclic loading measurements to assess the effects of the loading protocol along with the feedback control-based cyclic fatigue loading regimen. 
We used 25 Sprague-Dawley rats for this study, with 5 rats per group receiving either 500, 1,000, 2,000, 3,600, or 7,200 cycles of fatigue loads. The percentage differences between the pre- and post-cyclic loading measurements of the hysteresis, peak stress, and loading and unloading moduli were calculated. The results demonstrate that the system can induce varying degrees of damage to the Achilles tendon based on the number of loads applied. This system offers an innovative approach to apply quantified and physiological varying degrees of cyclic loads to the Achilles tendon for an in vivo model of fatigue-induced overuse tendon injury.

This protocol presents a testing system used to induce quantifiable and controlled fatigue injuries in a rat Achilles tendon for an in-vivo model of overuse-induced tendinopathy. The procedure consists of securing the rat's ankle to a joint actuator that performs passive ankle dorsiflexion with a custom-written MATLAB script.