High-resolution 3D volumetry versus conventional measuring techniques for the assessment of experimental lymphedema in the mouse hindlimb
Identifieur interne : 000974 ( Pmc/Curation ); précédent : 000973; suivant : 000975High-resolution 3D volumetry versus conventional measuring techniques for the assessment of experimental lymphedema in the mouse hindlimb
Auteurs : Florian S. Frueh [Allemagne, Suisse] ; Christina Körbel [Allemagne] ; Laura Gassert [Allemagne] ; Andreas Müller [Allemagne] ; Epameinondas Gousopoulos [Suisse] ; Nicole Lindenblatt [Suisse] ; Pietro Giovanoli [Suisse] ; Matthias W. Laschke [Allemagne] ; Michael D. Menger [Allemagne]Source :
- Scientific Reports [ 2045-2322 ] ; 2016.
Abstract
Secondary lymphedema is a common complication of cancer treatment characterized by chronic limb swelling with interstitial inflammation. The rodent hindlimb is a widely used model for the evaluation of novel lymphedema treatments. However, the assessment of limb volume in small animals is challenging. Recently, high-resolution three-dimensional (3D) imaging modalities have been introduced for rodent limb volumetry. In the present study we evaluated the validity of microcomputed tomography (μCT), magnetic resonance imaging (MRI) and ultrasound in comparison to conventional measuring techniques. For this purpose, acute lymphedema was induced in the mouse hindlimb by a modified popliteal lymphadenectomy. The 4-week course of this type of lymphedema was first assessed in 6 animals. In additional 12 animals, limb volumes were analyzed by μCT, 9.4 T MRI and 30 MHz ultrasound as well as by planimetry, circumferential length and paw thickness measurements. Interobserver correlation was high for all modalities, in particular for μCT analysis (r = 0.975, p < 0.001). Importantly, caliper-measured paw thickness correlated well with μCT (r = 0.861), MRI (r = 0.821) and ultrasound (r = 0.800). Because the assessment of paw thickness represents a time- and cost-effective approach, it may be ideally suited for the quantification of rodent hindlimb lymphedema.
Url:
DOI: 10.1038/srep34673
PubMed: 27698469
PubMed Central: 5048170
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Frueh</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Korbel, Christina" sort="Korbel, Christina" uniqKey="Korbel C" first="Christina" last="Körbel">Christina Körbel</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gassert, Laura" sort="Gassert, Laura" uniqKey="Gassert L" first="Laura" last="Gassert">Laura Gassert</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Muller, Andreas" sort="Muller, Andreas" uniqKey="Muller A" first="Andreas" last="Müller">Andreas Müller</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Clinic of Diagnostic and Interventional Radiology, Saarland University Medical Center</institution>, Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gousopoulos, Epameinondas" sort="Gousopoulos, Epameinondas" uniqKey="Gousopoulos E" first="Epameinondas" last="Gousopoulos">Epameinondas Gousopoulos</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology</institution>, ETH Zurich, Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lindenblatt, Nicole" sort="Lindenblatt, Nicole" uniqKey="Lindenblatt N" first="Nicole" last="Lindenblatt">Nicole Lindenblatt</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Giovanoli, Pietro" sort="Giovanoli, Pietro" uniqKey="Giovanoli P" first="Pietro" last="Giovanoli">Pietro Giovanoli</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Laschke, Matthias W" sort="Laschke, Matthias W" uniqKey="Laschke M" first="Matthias W." last="Laschke">Matthias W. 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Frueh</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Korbel, Christina" sort="Korbel, Christina" uniqKey="Korbel C" first="Christina" last="Körbel">Christina Körbel</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gassert, Laura" sort="Gassert, Laura" uniqKey="Gassert L" first="Laura" last="Gassert">Laura Gassert</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Muller, Andreas" sort="Muller, Andreas" uniqKey="Muller A" first="Andreas" last="Müller">Andreas Müller</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Clinic of Diagnostic and Interventional Radiology, Saarland University Medical Center</institution>, Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Gousopoulos, Epameinondas" sort="Gousopoulos, Epameinondas" uniqKey="Gousopoulos E" first="Epameinondas" last="Gousopoulos">Epameinondas Gousopoulos</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology</institution>, ETH Zurich, Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lindenblatt, Nicole" sort="Lindenblatt, Nicole" uniqKey="Lindenblatt N" first="Nicole" last="Lindenblatt">Nicole Lindenblatt</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Giovanoli, Pietro" sort="Giovanoli, Pietro" uniqKey="Giovanoli P" first="Pietro" last="Giovanoli">Pietro Giovanoli</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Laschke, Matthias W" sort="Laschke, Matthias W" uniqKey="Laschke M" first="Matthias W." last="Laschke">Matthias W. Laschke</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Menger, Michael D" sort="Menger, Michael D" uniqKey="Menger M" first="Michael D." last="Menger">Michael D. Menger</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Secondary lymphedema is a common complication of cancer treatment characterized by chronic limb swelling with interstitial inflammation. The rodent hindlimb is a widely used model for the evaluation of novel lymphedema treatments. However, the assessment of limb volume in small animals is challenging. Recently, high-resolution three-dimensional (3D) imaging modalities have been introduced for rodent limb volumetry. In the present study we evaluated the validity of microcomputed tomography (μCT), magnetic resonance imaging (MRI) and ultrasound in comparison to conventional measuring techniques. For this purpose, acute lymphedema was induced in the mouse hindlimb by a modified popliteal lymphadenectomy. The 4-week course of this type of lymphedema was first assessed in 6 animals. In additional 12 animals, limb volumes were analyzed by μCT, 9.4 T MRI and 30 MHz ultrasound as well as by planimetry, circumferential length and paw thickness measurements. Interobserver correlation was high for all modalities, in particular for μCT analysis (r = 0.975, p < 0.001). Importantly, caliper-measured paw thickness correlated well with μCT (r = 0.861), MRI (r = 0.821) and ultrasound (r = 0.800). 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">27698469</article-id><article-id pub-id-type="pmc">5048170</article-id><article-id pub-id-type="pii">srep34673</article-id><article-id pub-id-type="doi">10.1038/srep34673</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>High-resolution 3D volumetry versus conventional measuring techniques for the assessment of experimental lymphedema in the mouse hindlimb</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Frueh</surname><given-names>Florian S.</given-names></name><xref ref-type="corresp" rid="c1">a</xref><xref ref-type="aff" rid="a1">1</xref><xref ref-type="aff" rid="a2">2</xref><xref ref-type="author-notes" rid="n1">*</xref></contrib><contrib contrib-type="author"><name><surname>Körbel</surname><given-names>Christina</given-names></name><xref ref-type="aff" rid="a1">1</xref><xref ref-type="author-notes" rid="n1">*</xref></contrib><contrib contrib-type="author"><name><surname>Gassert</surname><given-names>Laura</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Müller</surname><given-names>Andreas</given-names></name><xref ref-type="aff" rid="a3">3</xref></contrib><contrib contrib-type="author"><name><surname>Gousopoulos</surname><given-names>Epameinondas</given-names></name><xref ref-type="aff" rid="a4">4</xref></contrib><contrib contrib-type="author"><name><surname>Lindenblatt</surname><given-names>Nicole</given-names></name><xref ref-type="aff" rid="a2">2</xref></contrib><contrib contrib-type="author"><name><surname>Giovanoli</surname><given-names>Pietro</given-names></name><xref ref-type="aff" rid="a2">2</xref></contrib><contrib contrib-type="author"><name><surname>Laschke</surname><given-names>Matthias W.</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Menger</surname><given-names>Michael D.</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><aff id="a1"><label>1</label><institution>Institute for Clinical and Experimental Surgery, Saarland University</institution>, 66421 Homburg/Saar,<country>Germany</country></aff><aff id="a2"><label>2</label><institution>Division of Plastic Surgery and Hand Surgery, University Hospital Zurich</institution>, 8091 Zurich,<country>Switzerland</country></aff><aff id="a3"><label>3</label><institution>Clinic of Diagnostic and Interventional Radiology, Saarland University Medical Center</institution>, Homburg/Saar,<country>Germany</country></aff><aff id="a4"><label>4</label><institution>Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology</institution>, ETH Zurich, Zurich,<country>Switzerland</country></aff></contrib-group><author-notes><corresp id="c1"><label>a</label><email>florian.frueh@uks.eu</email></corresp><fn id="n1"><label>*</label><p>These authors contributed equally to this work.</p></fn></author-notes><pub-date pub-type="epub"><day>04</day><month>10</month><year>2016</year></pub-date><pub-date pub-type="collection"><year>2016</year></pub-date><volume>6</volume><elocation-id>34673</elocation-id><history><date date-type="received"><day>14</day><month>07</month><year>2016</year></date><date date-type="accepted"><day>16</day><month>09</month><year>2016</year></date></history><permissions><copyright-statement>Copyright © 2016, The Author(s)</copyright-statement><copyright-year>2016</copyright-year><copyright-holder>The Author(s)</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link></license-p></license></permissions><abstract><p>Secondary lymphedema is a common complication of cancer treatment characterized by chronic limb swelling with interstitial inflammation. The rodent hindlimb is a widely used model for the evaluation of novel lymphedema treatments. However, the assessment of limb volume in small animals is challenging. Recently, high-resolution three-dimensional (3D) imaging modalities have been introduced for rodent limb volumetry. In the present study we evaluated the validity of microcomputed tomography (μCT), magnetic resonance imaging (MRI) and ultrasound in comparison to conventional measuring techniques. For this purpose, acute lymphedema was induced in the mouse hindlimb by a modified popliteal lymphadenectomy. The 4-week course of this type of lymphedema was first assessed in 6 animals. In additional 12 animals, limb volumes were analyzed by μCT, 9.4 T MRI and 30 MHz ultrasound as well as by planimetry, circumferential length and paw thickness measurements. Interobserver correlation was high for all modalities, in particular for μCT analysis (r = 0.975, p < 0.001). Importantly, caliper-measured paw thickness correlated well with μCT (r = 0.861), MRI (r = 0.821) and ultrasound (r = 0.800). Because the assessment of paw thickness represents a time- and cost-effective approach, it may be ideally suited for the quantification of rodent hindlimb lymphedema.</p></abstract></article-meta></front><floats-group><fig id="f1"><label>Figure 1</label><caption><title>Animal model.</title><p>(<bold>a–c</bold>) After methylene blue injection (<bold>a</bold>) the popliteal lymph node was visualized (<bold>b</bold>, asterisk). The afferent lymphatic vessels ran parallel to the iscial vein and were ligated (<bold>b</bold>, arrow). Subsequently, the popliteal fat pad including lymph nodes and efferent lymphatic vessels was resected (<bold>c</bold>). (<bold>d–f</bold>) Stereomicroscopic images illustrating paw edema with regression between day 3 (<bold>d</bold>) and day 10 (<bold>f</bold>). (<bold>g</bold>) Acute lymphedema in the mouse hindlimb over 28 days. Four experimental groups (n = 3 per group) were measured twice during the phase of postsurgical swelling (colored arrows) using different volumetric techniques. At the end of the experiment, there was still significant paw swelling (n = 6; mean ± SEM; *p < 0.05). (<bold>h,i</bold>) HE-stained paw cross-sections with increased dermal thickness 3 days (<bold>h</bold>) after lymph node dissection. Ten days after surgery (<bold>i</bold>), the paw volume had markedly decreased. Scales = 1 mm. (<bold>j,k</bold>) Inserts of (<bold>h</bold>,<bold>i)</bold>. Dermal lymphatic vessels were dilated on day 3 (<bold>j</bold>, arrowheads), but exhibited normal configuration on day 10 (<bold>k</bold>, arrowheads) as shown by means of immunohistochemical staining with LYVE-1. Scales = 140 μm.</p></caption><graphic xlink:href="srep34673-f1"></graphic></fig><fig id="f2"><label>Figure 2</label><caption><title>µCT, MRI and hrUS for hindlimb volumetry.</title><p>(<bold>a–l</bold>) Coronal (<bold>a–f</bold>) and axial (<bold>g–l</bold>) hindlimb images of μCT (<bold>a,b,g</bold>,<bold>h</bold>), MRI (<bold>c,d,i</bold>,<bold>j</bold>) and hrUS (<bold>e,f,k</bold>,<bold>l</bold>) 3 days after popliteal lymphadenectomy. In T2-weighted MR images, the thickened and edematous tissue was characterized by epifascial hyperintensity (<bold>c</bold>, arrowheads). In contrast, 30 MHz hrUS objectified lymphedema as a hypoechoic layer (<bold>e</bold>, arrowheads). (<bold>e</bold>) Arrow = gluteal fold, which was used as a landmark for volume calculation. In axial images, tibia (<bold>g–l</bold>, white arrowhead) and fibula (<bold>g–j</bold>, black arrowheads) can be reliably identified. Note the dorsal acoustic attenuation in hrUS (<bold>k</bold>,<bold>l</bold>, asterisk). LE = lymphedema; scales (<bold>a–f)</bold> = 4.5 mm; (<bold>g–l)</bold> = 3 mm.</p></caption><graphic xlink:href="srep34673-f2"></graphic></fig><fig id="f3"><label>Figure 3</label><caption><title>Interobserver variability of hindlimb volumetry modalities.</title><p>(<bold>a–l</bold>) μCT showed the highest correlation between two observers (<bold>a</bold>) and a low measurement variability (<bold>b</bold>) compared to other modalities as assessed by linear correlation (<bold>a,c,e,g,i</bold>,<bold>k</bold>) and Bland-Altman analyses (<bold>b,d,f,h,j</bold>,<bold>l</bold>). Despite low interobserver variability, MRI, hrUS, planimetry and MRI-measured circumference were associated with higher measurement variations (<bold>c–j</bold>). Dashed line = mean difference between observers; dotted line = double standard deviation. n = 48 for μCT, MRI, hrUS, MRI circumference and caliper; n = 42 for planimetry.</p></caption><graphic xlink:href="srep34673-f3"></graphic></fig><fig id="f4"><label>Figure 4</label><caption><title>Correlation between different 3D volumetry modalities.</title><p>(<bold>a–f</bold>) Linear correlations (<bold>a,c</bold>,<bold>e</bold>) and Bland-Altman plots (<bold>b,d</bold>,<bold>f</bold>) of 3D volumetry using the gluteal fold as landmark. Hindlimb volumes are illustrated in grey scale with non-operated controls (white circles) and operated limbs (day 3–10: black to bright grey circles). The volumes calculated with the gluteal fold landmark correlated poorly among the 3D modalities and led to a random distribution of the volumes without the expected grouping into control and operated hindlimb volumes. (<bold>g–l</bold>) After standardized measuring with the distal TF-joint landmark, 3D volumetry exhibited good correlations (<bold>g,i</bold>,<bold>k</bold>). However, the higher the hindlimb volumes, the higher the measurement variability was as shown in Bland-Altman analyses (<bold>h,j</bold>,<bold>l</bold>; black circles). Dashed line = mean difference between volume measurements; dotted line = double standard deviation; n = 48.</p></caption><graphic xlink:href="srep34673-f4"></graphic></fig><fig id="f5"><label>Figure 5</label><caption><title>The distal TF-joint as landmark for standardized 3D limb volumetry.</title><p>(<bold>a–e</bold>) The distal TF-joint was located in the distal third of the mouse hindlimb, as shown in an illustration (<bold>a</bold>) and 3D reconstruction of the hindlimb skeleton (<bold>b</bold>). It could be reliably localized in axial images of 3D modalities (<bold>c,d</bold>,<bold>e</bold>). Proximal to the TF-joint (<bold>c</bold>), both tibia (white arrowhead) and fibula (black arrowhead) were detectable. More distally, the two bones articulated (<bold>d</bold>, arrow). The level of the TF-joint was used for MRI-based calculations of limb circumference. (<bold>a</bold>) was drawn by Carol De Simio (University Hospital Zurich).</p></caption><graphic xlink:href="srep34673-f5"></graphic></fig><fig id="f6"><label>Figure 6</label><caption><title>3D volume and circumference measurements with MRI.</title><p>(<bold>a–c</bold>) The two types of measurement correlated (<bold>a</bold>) but 3D volumetry revealed markedly higher limb ratios, as depicted by Bland-Altman analysis (<bold>b</bold>). Dashed line = mean difference between ratios; dotted line = double standard deviation; n = 48; day 3-10 = black to bright grey circles; non-operated control limbs = white circles. <bold>(c)</bold> Limb ratios based on 3D volumes were consistently higher throughout the course of the study (3D volume: black circles; circumference: white circles; n = 3 for each time point; mean ± SEM; *p < 0.05).</p></caption><graphic xlink:href="srep34673-f6"></graphic></fig><fig id="f7"><label>Figure 7</label><caption><title>Caliper-measured paw thickness versus other hindlimb volumetry modalities.</title><p>(<bold>a–j</bold>) Linear correlations (<bold>a,c,e,g</bold>,<bold>i</bold>) and Bland-Altman plots (<bold>b,d,f,h</bold>,<bold>j</bold>) comparing caliper-measured paw thickness with other hindlimb volumetry modalities. The caliper correlated well with 3D volumetry (<bold>a–f</bold>) but no or low correlation with planimetry (<bold>g,h</bold>) and circumferential length (<bold>i,j</bold>) was recorded. Importantly, hindlimb ratios based on paw thickness were higher than those calculated with the other modalities (n = 48 for μCT, MRI, hrUS, MRI circumference and caliper; n = 42 for planimetry; day 3–10 = black to bright grey circles; non-operated control limbs = white circles; dashed line = mean difference between ratios; dotted line = double standard deviation).</p></caption><graphic xlink:href="srep34673-f7"></graphic></fig><table-wrap position="float" id="t1"><label>Table 1</label><caption><title>Features of different techniques for rodent hindlimb volumetry.</title></caption><table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col><col align="center"></col><col align="center"></col><col align="center"></col><col align="center"></col></colgroup><thead valign="bottom"><tr><th align="left" valign="top" charoff="50">Modality</th><th align="center" valign="top" charoff="50">Costs</th><th align="center" valign="top" charoff="50">Examination time (min ± SD)</th><th align="center" valign="top" charoff="50">Anesthesia required?</th><th align="center" valign="top" charoff="50">Lymphatic imaging possible?</th></tr></thead><tbody valign="top"><tr><td align="left" valign="top" charoff="50">μCT</td><td align="center" valign="top" charoff="50">+ + +</td><td align="center" valign="top" charoff="50">14.6 ± 2.2</td><td align="center" valign="top" charoff="50">yes</td><td align="center" valign="top" charoff="50">yes</td></tr><tr><td align="left" valign="top" charoff="50">MRI</td><td align="center" valign="top" charoff="50">+ + +</td><td align="center" valign="top" charoff="50">34.9 ± 2.8</td><td align="center" valign="top" charoff="50">yes</td><td align="center" valign="top" charoff="50">yes</td></tr><tr><td align="left" valign="top" charoff="50">hrUS</td><td align="center" valign="top" charoff="50">+ + +</td><td align="center" valign="top" charoff="50">4.9 ± 0.9</td><td align="center" valign="top" charoff="50">yes</td><td align="center" valign="top" charoff="50">no (yes)</td></tr><tr><td align="left" valign="top" charoff="50">Planimetry</td><td align="center" valign="top" charoff="50">+ +</td><td align="center" valign="top" charoff="50">1.0 ± 0.3</td><td align="center" valign="top" charoff="50">yes</td><td align="center" valign="top" charoff="50">no</td></tr><tr><td align="left" valign="top" charoff="50">Circumference</td><td align="center" valign="top" charoff="50">+</td><td align="center" valign="top" charoff="50">not assessed</td><td align="center" valign="top" charoff="50">no</td><td align="center" valign="top" charoff="50">no</td></tr><tr><td align="left" valign="top" charoff="50">Caliper</td><td align="center" valign="top" charoff="50">+</td><td align="center" valign="top" charoff="50">1.2 ± 0.2</td><td align="center" valign="top" charoff="50">no</td><td align="center" valign="top" charoff="50">no</td></tr></tbody></table><table-wrap-foot><fn id="t1-fn1"><p>3D techniques are associated with high costs, longer examination times (i.e. time span from anesthetic induction to end of imaging) and always require anesthesia. They bear the advantage of additional lymphatic imaging using lymph-affine contrast agents. In contrast, conventional techniques are time- and cost-efficient methods to assess surrogate parameters for rodent hindlimb volumes. For maximal accuracy, hindlimb planimetry should be performed under anesthesia as well.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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