Name: biotribological testing and analysis of articular cartilage sliding against metal for implants
Uploaded: 2020-05-14T15:00:00
Description: Watch this Scientific Journal Video about Biotribological Testing and Analysis of Articular Cartilage Sliding against Metal for Implants at JoVE.com

这项研究由Né Forschungs-und Bildungsges.m.b.H.和下奥地利省政府通过生命科学电话（项目ID：LSC15-019）和奥地利彗星方案（项目K2 XTribology，第849109号赠款）。

1. 金属钢瓶的准备
<ol><li>分析圆柱形钴-铬-硅（CoCrMo）棒，使用扫描电子显微镜（SEM）和每个制造商的协议能量分散X射线光谱，以确认所提供的值，符合手术植入物的化学成分标准规范。 注：用于此实验的CoCrMo合金元素成分为65%Co、28%Cr、5%Mo和2%其他。</li>
	<li>用碳化硅研磨纸湿磨样品，从粒度为 500 开始。使用研磨纸，增加顺序高达 4000 的颗粒大小。</li>
	<li>用 3 μm 和 1 μm 糊进行抛光，以达?...

<ol>
	<li>Zengerink, M., Struijs, P. A. A. A., Tol, J. L., van Dijk, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+osteochondral+lesions+of+the+talus:+a+systematic+review.">Treatment of osteochondral lesions of the talus: a systematic review.</a> Knee Surgery, Sports Traumatology, Arthroscopy. 18 (2), 238-246 (2009).</li><li>Aurich, 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=Behandlung+osteochondraler+L&#228;sionen+des+Sprunggelenks:+Empfehlungen+der+Arbeitsgemeinschaft+Klinische+Geweberegeneration+der+DGOU.">Behandlung osteochondraler L&#228;sionen des Sprunggelenks: Empfehlungen der Arbeitsgemeinschaft Klinische Geweberegeneration der DGOU.</a> Zeitschrift fur Orthopadie und Unfallchirurgie. 155 (1), 92-99 (2017).</li><li>Van Bergen, C. J. A., Zengerink, M., Blankevoort, L., Van Sterkenburg, M. N., Van Oldenrijk, J., Van Dijk, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+metallic+implantation+technique+for+osteochondral+defects+of+the+medial+talar+dome.">Novel metallic implantation technique for osteochondral defects of the medial talar dome.</a> Acta Orthopaedica. 81 (4), 495-502 (2010).</li><li>Sweet, S. J., Takara, T., Ho, L., Tibone, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primary+Partial+Humeral+Head+Resurfacing.">Primary Partial Humeral Head Resurfacing.</a> The American Journal of Sports Medicine. 43 (3), 579-587 (2015).</li><li>Becher, C., 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=Minimum+5-year+results+of+focal+articular+prosthetic+resurfacing+for+the+treatment+of+full-thickness+articular+cartilage+defects+in+the+knee.">Minimum 5-year results of focal articular prosthetic resurfacing for the treatment of full-thickness articular cartilage defects in the knee.</a> Archives of Orthopaedic and Trauma Surgery. 131 (8), 1135-1143 (2011).</li><li>Lea, M. A., Barkatali, B., Porter, M. L., Board, T. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteochondral+Lesion+of+the+Hip+Treated+with+Partial+Femoral+Head+Resurfacing.+Case+Report+and+Six-Year+Follow-up.">Osteochondral Lesion of the Hip Treated with Partial Femoral Head Resurfacing. Case Report and Six-Year Follow-up.</a> HIP International. 24 (4), 417-420 (2018).</li><li>Becher, C., Huber, R., Thermann, H., Paessler, H. H., Skrbensky, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+a+contoured+articular+prosthetic+device+on+tibiofemoral+peak+contact+pressure:+a+biomechanical+study.">Effects of a contoured articular prosthetic device on tibiofemoral peak contact pressure: a biomechanical study.</a> Knee Surgery, Sports Traumatology, Arthroscopy. 16 (1), 56-63 (2007).</li><li>Malahias, M. -. A., Chytas, D., Thorey, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+clinical+outcome+of+the+different+HemiCAP+and+UniCAP+knee+implants:+A+systematic+and+comprehensive+review.">The clinical outcome of the different HemiCAP and UniCAP knee implants: A systematic and comprehensive review.</a> Orthopedic Reviews. 10 (2), (2018).</li><li>Dhollander, A. A. 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=The+use+of+a+prosthetic+inlay+resurfacing+as+a+salvage+procedure+for+a+failed+cartilage+repair.">The use of a prosthetic inlay resurfacing as a salvage procedure for a failed cartilage repair.</a> Knee Surgery, Sports Traumatology. 23 (8), 2208-2212 (2014).</li><li>Van Bergen, C. J. A. A., van Eekeren, I. C. M. M., Reilingh, M. L., Sierevelt, I. N., van Dijk, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+osteochondral+defects+of+the+talus+with+a+metal+resurfacing+inlay+implant+after+failed+previous+surgery.">Treatment of osteochondral defects of the talus with a metal resurfacing inlay implant after failed previous surgery.</a> Bone and Joint Journal. 95 (12), 1650-1655 (2013).</li><li>Kim, Y. S. Y. -. H. H. Y. -. S., Kim, Y. S. Y. -. H. H. Y. -. S., Hwang, K. -. T. T., Choi, I. -. Y. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cartilage+degeneration+and+joint+motion+of+bipolar+hemiarthroplasty.">The cartilage degeneration and joint motion of bipolar hemiarthroplasty.</a> International Orthopaedics. 36 (10), 2015-2020 (2012).</li><li>Moon, K. H., 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=Degeneration+of+Acetabular+Articular+Cartilage+to+Bipolar+Hemiarthroplasty.">Degeneration of Acetabular Articular Cartilage to Bipolar Hemiarthroplasty.</a> Yonsei Medical Journal. 49 (5), 716-719 (2008).</li><li>Wimmer, M. A., Pacione, C., Laurent, M. P., Chubinskaya, S. <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+wear+testing+of+living+cartilage+articulating+against+alumina.">In vitro wear testing of living cartilage articulating against alumina.</a> Journal of Orthopaedic Research. , (2016).</li><li>Bowland, P., Ingham, E., Fisher, J., Jennings, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+geometry+tribological+study+of+osteochondral+graft+implantation+in+the+knee.">Simple geometry tribological study of osteochondral graft implantation in the knee.</a> Proceedings of the Institution of Mechanical Engineers, Part H: Journal of Engineering in Medicine. 232 (3), 249-256 (2018).</li><li>Bowland, P., Ingham, E., Fisher, J., Jennings, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+preclinical+natural+porcine+knee+simulation+model+for+the+tribological+assessment+of+osteochondral+grafts+in+vitro.">Development of a preclinical natural porcine knee simulation model for the tribological assessment of osteochondral grafts in vitro.</a> Journal of Biomechanics. 77, 91-98 (2018).</li><li>Trevino, R. L., 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=Establishing+a+live+cartilage-on-cartilage+interface+for+tribological+testing.">Establishing a live cartilage-on-cartilage interface for tribological testing.</a> Biotribology. 9, 1-11 (2017).</li><li>Oungoulian, S. R., 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=Wear+and+damage+of+articular+cartilage+with+friction+against+orthopedic+implant+materials.">Wear and damage of articular cartilage with friction against orthopedic implant materials.</a> Journal of Biomechanics. 48 (10), 1957-1964 (2015).</li><li>Stotter, C., 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=Effects+of+Loading+Conditions+on+Articular+Cartilage+in+a+Metal-on-Cartilage+Pairing.">Effects of Loading Conditions on Articular Cartilage in a Metal-on-Cartilage Pairing.</a> Journal of Orthopaedic Research. 37 (12), 2531-2539 (2019).</li><li>Becher, C., Huber, R., Thermann, H., Tibesku, C. O., von Skrbensky, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tibiofemoral+contact+mechanics+with+a+femoral+resurfacing+prosthesis+and+a+non-functional+meniscus.">Tibiofemoral contact mechanics with a femoral resurfacing prosthesis and a non-functional meniscus.</a> Clinical biomechanics. 24 (8), 648-654 (2009).</li><li>Temple, D. K., Cederlund, A. A., Lawless, B. M., Aspden, R. M., Espino, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viscoelastic+properties+of+human+and+bovine+articular+cartilage:+a+comparison+of+frequency-dependent+trends.">Viscoelastic properties of human and bovine articular cartilage: a comparison of frequency-dependent trends.</a> BMC Musculoskeletal Disorders. , 1-8 (2016).</li><li>Caligaris, M., Ateshian, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+sustained+interstitial+fluid+pressurization+under+migrating+contact+area,+and+boundary+lubrication+by+synovial+fluid,+on+cartilage+friction.">Effects of sustained interstitial fluid pressurization under migrating contact area, and boundary lubrication by synovial fluid, on cartilage friction.</a> Osteoarthritis and Cartilage. 16 (10), 1220-1227 (2008).</li><li>Burris, D. L., Ramsey, L., Graham, B. T., Price, C., Moore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+Sliding+and+Hydrodynamics+Contribute+to+Articular+Cartilage+Fluid+and+Lubrication+Recovery.">How Sliding and Hydrodynamics Contribute to Articular Cartilage Fluid and Lubrication Recovery.</a> Tribology Letters. 67 (2), 1-10 (2019).</li><li>Mamat, N., Nor, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Numerical+measurement+of+contact+pressure+in+the+tibiofemoral+joint+during+gait.">Numerical measurement of contact pressure in the tibiofemoral joint during gait.</a> International Conference on Biomedical Engineering (ICoBE). , 27-28 (2012).</li><li>Manda, K., Ryd, L., Eriksson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Finite+element+simulations+of+a+focal+knee+resurfacing+implant+applied+to+localized+cartilage+defects+in+a+sheep+model.">Finite element simulations of a focal knee resurfacing implant applied to localized cartilage defects in a sheep model.</a> Journal of Biomechanics. 44 (5), 794-801 (2011).</li></ol>

聚焦金属植入物是骨质疏松缺陷的抢救程序，特别是在中年患者和初级治疗失败后。虽然临床研究表明有希望的短期结果，一个观察到的并发症是损害对立的，本地软骨10。尸体和生物力学研究表明，适当的植入与平坦或轻微凹陷定位保持自然接触压力19。摩擦实验提供了在体外测试各种软骨配对的可能性。在这种情况下，装载条件、润滑、材料配对和持续时间...

骨质疏松缺陷的治疗要求很高，在许多情况下需要手术。对于中年患者的聚焦骨质病变，焦点金属植入物是一个可行的选择，尤其是在原发治疗失败之后，如骨髓刺激（BMS）或自体软骨细胞植入（ACI）1。部分表面置换可视为挽救程序，可减轻疼痛，改善运动范围2。这些植入物通常由CoCrMo合金组成，具有不同的尺寸和偏移配置，以匹配正常的解剖结构3。虽然最初开发为膝盖的内侧股骨孔的缺陷，这种植入物现在可用，并用于臀部，脚踝，肩膀和肘部4，5，6。为了获得满意的结果，评估相对软骨的机械关节对齐和状况至关重要。此外，正确的植入没有突起植入物已被证明是基本的7。
临床研究显示，在减少疼痛及改善中年病人功能方面，有优良的短期效果。与同体植入相比，焦金属植入物允许早期承重。然而，对立的关节软骨显示加速磨损在相当多的病人9，10。因此，即使放置适当，在许多情况下，原生软骨的退化似乎也不可避免，而基本机制仍然不清楚。类似的退行性变化已经观察到后，双相体体成形术的臀部11和增加与活动和加载12。
摩擦实验提供了在体外研究这种配对和模拟生理条件下发生的不同载荷情况的可能性。骨质针的使用提供了一个简单的几何模型，以研究关节软骨滑对原生软骨或任何植入材料14的摩擦学， 并可以进一步用于整个关节模拟模型15。软骨上的金属配对显示软骨磨损加速，细胞外基质中断，和减少细胞生存能力在表面区域相比软骨对软骨配对16。软骨的损伤主要发生在浅层和中间区域17之间的分层形式。然而，导致软骨退化的机制尚未完全了解。该协议对关节软骨的生物合成活性进行了综合分析。通过确定代谢活性和代谢基因的基因表达水平，可以确定软骨分解的早期适应症。体外摩擦学实验的优点是可以调整载荷参数以模拟各种载荷条件。
因此，以下协议适合模拟软骨上的金属配对，代表实验性半人成形模型。

<table border="0" cellpadding="0" cellspacing="0" style="width:773px;" width="772">
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	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:297px;">Amphotericin B</td>
			<td style="width:191px;">Sigma&#8208;Aldrich Chemie GmbH</td>
			<td style="width:119px;">A-2942-100ML</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">buffered formaldehyde solution 4%</td>
			<td style="width:191px;">VWR</td>
			<td style="width:119px;">97131000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cell Proliferation Kit II (XTT)</td>
			<td>Roche Diagnostics</td>
			<td style="width:119px;">11465015001</td>
			<td>XTT-based ex vivo toxicology assay</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">CoCrMo raw material</td>
			<td style="width:191px;">Acnis International</td>
			<td style="width:119px;"></td>
			<td>CoCrMo rods 6mm in diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CryoStar NX70 Cryostat</td>
			<td>Thermo Fischer Scientific</td>
			<td style="width:119px;"></td>
			<td>cryosectioning device</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">dimethyl sulfoxide (DMSO)</td>
			<td style="width:191px;">Sidma-Aldrich Chemie</td>
			<td style="width:119px;">D 2438-10ML</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Dulbecco&#8217;s modified Eagle&#8217;s medium</td>
			<td style="width:191px;">Sigma&#8208;Aldrich Chemie GmbH</td>
			<td style="width:119px;"></td>
			<td>medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">fetal bovine serum</td>
			<td style="width:191px;">Gibco</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Hyaluronic acid</td>
			<td style="width:191px;">Anika Therapeutics Inc.</td>
			<td style="width:119px;"></td>
			<td>component of lubricating solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">iCycler</td>
			<td>BioRad</td>
			<td style="width:119px;"></td>
			<td>thermal cycler</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Leica microscope DM&#8208;1000</td>
			<td>Leica</td>
			<td style="width:119px;"></td>
			<td>microscope for histology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">LightCycler 480 Sealing Foil</td>
			<td style="width:191px;">Roche Diagnostics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">LightCycler 96</td>
			<td>Roche Diagnostics</td>
			<td style="width:119px;"></td>
			<td>thermal cycler for PCR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MagNA Lyser Green Beads</td>
			<td>Roche Diagnostics</td>
			<td style="width:119px;">3358941001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Osteochondral Autograft Transfer System (OATS)</td>
			<td style="width:191px;">Arthrex Inc.</td>
			<td style="width:119px;"></td>
			<td>cutting tube for harvesting osteochondral cylinders</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">osteosoft</td>
			<td style="width:191px;">Merck</td>
			<td style="width:119px;">1017279010</td>
			<td>decalcifier-solution</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Penicillin /Streptomycin</td>
			<td style="width:191px;">Sigma&#8208;Aldrich Chemie GmbH</td>
			<td style="width:119px;">P4333-100ML</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:297px;">phosphate&#8208;buffered saline</td>
			<td style="width:191px;">Sigma&#8208;Aldrich Chemie GmbH</td>
			<td style="width:119px;"></td>
			<td>PBS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Prescale Low Pressure</td>
			<td style="width:191px;">Fujifilm</td>
			<td style="width:119px;"></td>
			<td>pressure indicating film</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RNeasy Fibrous Tissue Kit</td>
			<td style="width:191px;">QIAGEN</td>
			<td style="width:119px;">74404</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Synergy 2</td>
			<td>BioTek Instruments</td>
			<td style="width:119px;"></td>
			<td>plate reader</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Tetra&#8208;Falex MUST</td>
			<td style="width:191px;">Falex Tribology</td>
			<td style="width:119px;"></td>
			<td>Tribometer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tissue&#8208; Tek O.C.T.</td>
			<td style="width:191px;">SAKURA</td>
			<td style="width:119px;">4583</td>
			<td>embedding formulation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transcriptor First Strand cDNA Synthesis Kit</td>
			<td>Roche Diagnostics</td>
			<td style="width:119px;">40897030001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#946;-mercaptoethanol</td>
			<td style="width:191px;">Sidma-Aldrich Chemie</td>
			<td style="width:119px;">M3148</td>
			<td></td>
		</tr>
	</tbody>
</table>

biotribological testing and analysis of articular cartilage sliding against metal for implants

中年患者的骨质疏松缺陷可能用焦点金属植入物进行治疗。首先开发为膝关节的缺陷，植入物现在可用于肩部，臀部，脚踝和第一个骨质性关节。在提供疼痛减轻和临床改善的同时，许多患者观察到对立软骨的渐进性退行性变化。导致这种损害的机制尚未完全了解。该协议描述了一个摩擦实验，以模拟软骨上的金属配对和关节软骨的综合分析。金属植入物材料测试牛骨柱作为人体关节软骨的模型。通过应用不同的负载和滑动速度，可以模拟生理载荷条件。为了全面分析关节软骨的影响，本协议描述了组织学、代谢活性和基因表达分析。摩擦学测试的主要优点是，可以自由调节载荷参数，以模拟体内条件。此外，不同的测试解决方案可用于研究润滑或亲炎剂的影响。通过使用软骨特异性基因和猫科动物基因的基因表达分析，可以检测到关节软骨细胞代谢的早期变化，以回应机械负荷。

必须使用压力测量膜确认接触区域和接触压力（图1）。生理载荷条件可以通过与所定义的接触压力的参考印迹进行比较来确认。在测试过程中，摩擦系数不断被监测。对于迁移接触区，低摩擦系数可保持至少 1 小时（图 2）。使用萨夫兰宁O染色的细胞外基质组成和结构可以确定（图3）。萨夫拉宁 O 染色的强度与蛋白细胞素含量?...

Watch this Scientific Journal Video about Biotribological Testing and Analysis of Articular Cartilage Sliding against Metal for Implants at JoVE.com

Biotribological Testing and Analysis of Articular Cartilage Sliding against Metal for Implants

该协议描述了骨质体油柱在金属植入物材料上滑动的制备、生物摩擦测试和分析。该协议中包含的结果指标是代谢活性、基因表达和组织学。

植入物关节软骨滑向金属的生物摩擦学测试与分析

Osteochondral defects in middle-aged patients might be treated with focal metallic implants. First developed for defects in the knee joint, implants are ...

我们的协议使得测试金属对关节软骨的滑动成为可能，从而研究焦距植入物或血液成形术对特定软骨细胞的生物合成活性的影响，该技术的主要优点是，它提供了对软骨配对中机械负荷的摩擦特性和生物效应的全面洞察。这项技术可能称赞手术后的临床发现，如植入焦金属植入物，以治疗膝盖的骨质缺陷或臀部的脑骨成形术。演示这个程序的将是克里斯托弗·鲍尔，一个来自我在多瑙河大学实验室的博士后。使用市售往复式摩擦计，带钢瓶，具有板配置、垂直装载功能以及可调负载和滑动速度。此外，在润滑溶液中执行测试需要液体电池。将骨质松缸固定到底部样品支架上，标记与滑动方向对齐。首先使用压力测量膜确定软骨系统钴铬硅中的接触压力。将压力测量膜放在界面上，并施加 30 秒的静态负载以确定初始接触压力、接触尺寸和形状。将钴铬硅气瓶安装到上部称重传感器上。将测试溶液加入液体电池，使骨质松体油缸淹没并覆盖金属软骨滑动界面。设置测试参数，如正常力冲程和滑动速度所述，在整个测试过程中将保持不变。将金属油缸与浸入润滑溶液中的关节软骨开始相互滑动。在整个实验中监测摩擦系数，终止实验。欲望测试期后，从样品架中取出骨质松体塞，用PBS冲洗，并将其储存在介质中，直到进一步进行生物分析。在测试期间，将控制样品和测试解决方案保持在室温下，并结合暴露于机械负载的样品进行分析。放置，一个24井板在刻度上，零。用 PBS 冲洗骨质疏松塞，并放在培养皿中。然后使用手术刀将软骨从移植物中切成两部分，将软骨分成两部分，使接触区域均匀地分布在软骨碎片上，将一半切成大约一毫米的立方体。使用后半部分进行基因表达分析，将切碎的软骨转移到24井板的一个井中，并确定每个样品的组织重量重复这个过程，并将一毫升的生长介质添加到板的每个井中。将500微升的XTT溶液加入到每井中，然后混合在37摄氏度和5%的二氧化碳下孵育，孵育后4小时取出上经剂并将其转移到5毫升管中。通过将 500 微升 DMSO 添加到每个井的软骨组织中，在室温下连续搅拌一小时，去除 DMSO 溶液，然后用先前收集的 XTT 溶液将其拉拔，从而提取四分之一糖。将每个样品的 100 微升三分之一转移到 96 井板中。使用板读卡器测量波长为 492 纳米的吸光度和 690 纳米的参考波长。为了分离RNA薄荷，从骨质疏松插入小块获得的软骨组织的后半部分将组织转移到含有陶瓷珠和300微升许可缓冲液的管中。使用 1%β甲醇使用商用乳酸酸酯，将应用 6，500 RPM 的组织均质化，四次应用 20 秒，每次运行后 20 分钟的冷却阶段。在每个样品中加入20微升蛋白酶K和580微升RNA无水，并在55摄氏度下孵育30分钟。将样品在10，000倍G下离心3分钟，然后将上流液转移到1.5毫升管中。将0.5卷90%乙醇加入每个管中，然后混合，然后将700微升样品转移到两毫升收集管中的RNA结合柱中，并在8000次G下离心15秒。丢弃流经，对乳酸盐的其余部分重复离心步骤。向列中添加 350 微升的缓冲 RW RW。在 8，000 次 G 下离心 15 秒，然后丢弃流经。混合 10 微升 DNA 库存溶液和 70 微升缓冲 RDD。将溶液加入RNA纯化膜，在室温下孵育15分钟，然后向柱中加入350微升缓冲鲁液。在8000次G下离心15秒，丢弃流式将500微升缓冲RPE添加到RNA纯化柱，并在8时离心。 000 次 G 15 秒丢弃流经，再向 RNA 纯化柱中添加 500 微升缓冲 RPE，然后在 8，000 次 G 下离心，两分钟将该柱放在新的 1.5 毫升收集管中，并添加 30 微升RNA 释放水在 8，000 次 G 下离心管一分钟，用商业试剂盒解冻和混合所有区域来表示RNA合成的 cDNA，加入RNA样本，执行文本手稿中描述的反应和热循环器。在 96 孔板的每个孔中加入 9 微升的 RTQ PCR 主混合物，并在 4 摄氏度下将 PCR 板与天花板油一起发光，并在 4 摄氏度下将其在 877 倍 G 下离心 10 分钟，执行 RTQ PCR 和精密热循环器。根据手稿指示。在测试金属软骨界面的接触区域和接触压力之前，必须使用压力测量膜进行确认。然后，可以通过将获得的印迹与定义的接触的参考推论进行比较来确认生理载荷条件。细胞外基质组成和结构可以通过藏红花和O染色确定。藏红花和 O 染色的强度与蛋白蛋白细胞含量成正比。在关节表面，蛋白糖含量变化，但应均匀地在整个组织部分的基线样品，显示提取糖基糖，可以通过机械加载抵消。牛关节脊椎细胞的代谢活性与收获地点无关，但随着机械载荷的增加，这种代谢活性也大为增加。软骨特异性基因的基因表达水平随着生理负荷条件而增加。而代谢基因是向上调节与固定接触区域。按照此过程，可以执行额外的分析，包括软骨网产品的测定和使用扫描电子显微镜对关节表面的分析，此外，我们尝试在各种摩擦学配对中优化关节软骨的润滑。

Research

JoVE Journal

Medicine

A Novel Application of Musculoskeletal Ultrasound Imaging

肌肉骨骼超声成像的一种新应用

Ultrasound is an attractive modality for imaging muscle and tendon  motion during dynamic tasks and can provide a complementary methodological  approach ...

科研

医学

Techniques for Processing Eyes Implanted with a Retinal Prosthesis for Localized Histopathological Analysis: Part 2 Epiretinal Implants with Retinal Tacks

为处理眼植入视网膜假体的局部病理组织学分析技术：第2部分视网膜前植入视网膜大头钉

Retinal prostheses for the treatment of certain forms of blindness are gaining traction in clinical trials around the world with commercial devices ...

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

近端股骨尸制备基于CT的断裂强度测试和定量的有限元分析

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

Oral Biofilm Formation on Different Materials for Dental Implants

牙种植体不同材料的口腔生物膜形成

Dental implants and their prosthetic components are prone to bacterial colonization and biofilm formation. The use of materials that provides low ...

Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

金属异物在儿童体内摄取的手持金属探测器筛选

Coins are the most common ingested metallic foreign bodies among children. The goal of this protocol is to assess the accuracy and feasibility of using a ...

Hemocompatibility Testing of Blood-Contacting Implants in a Flow Loop Model Mimicking Human Blood Flow

流循环模型中血液接触植入物的血液相容性测试模拟人类血流

The growing use of medical devices (e.g., vascular grafts, stents, and cardiac catheters) for temporary or permanent purposes that remain in the ...

Novel Percutaneous Approach for Deployment of 3D Printed Coronary Stenosis Implants in Swine Models of Ischemic Heart Disease

在缺血性心脏病的猪模型中部署3D打印冠状动脉狭窄植入物的新颖的皮下方法

Minimally invasive methods for creating models of focal coronary narrowing in large animals are challenging. Rapid prototyping using three-dimensionally ...

3D Planning and Printing of Patient Specific Implants for Reconstruction of Bony Defects

患者特异性植入物重建的三维规划与打印

We are in the midst of the 3D era in most aspects of life, and especially in medicine. The surgical discipline is one of the major players in the medical ...

Treatment of Facial Deformities using 3D Planning and Printing of Patient-Specific Implants

使用患者特定植入物的 3D 规划和打印治疗面部畸形

Technological advancements in surgical planning and patient-specific implants are constantly evolving. One can either adopt the technology to achieve ...