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Quantification of cell density in rat Achilles tendon: development and application of a new method

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@article{e0df03ff23004e029bef8b27d42f8631,
title = "Quantification of cell density in rat Achilles tendon: development and application of a new method",
abstract = "Increased tendon cell nuclei density (TCND) has been proposed to induce tendon mechanical adaptations. However, it is unknown whether TCND is increased in tendon tissue after mechanical loading and whether such an increase can be quantified in a reliable manner. The aim of this study was to develop a reliable method for quantification of TCND and to investigate potential changes in TCND in rat Achilles tendons in response to 12 weeks of running. Eight adult male Sprague-Dawley rats ran (RUN) on a treadmill with 10° incline, 1 h/day, 5 days/wk (17-20 m/min) for 12 weeks (which improved tendon mechanical properties) and were compared with 11 control rats (SED). Tissue-Tek-embedded cryosections (10 µm) from the mid region of the Achilles tendon were cut longitudinally on a cryostat. Sections were stained with alcian blue and picrosirius red. One blinded investigator counted the number of tendon cell nuclei 2-3 times in three separate regions of the mid longitudinal tendon sections with fields of 390 μm × 280 μm. Unpaired t tests were used for the statistical analysis (mean ± SE). Typical Error {\%} for replicate counts was 5.5 and 14 {\%} coefficient of variation for the three regions. There was no difference in TCND between running rats versus control rats (nuclei per image (≈105 μm2): RUN, 152 ± 9; SED, 146 ± 8, p = 0.642). This new method provided reproducible quantification of TCND. There was no difference in TCND despite improvements in tendon mechanics, which suggests that cell number is not a major cause for altered tendon mechanical properties with loading.",
keywords = "Achilles Tendon, Animals, Cell Count, Cell Nucleus, Male, Rats, Rats, Sprague-Dawley, Tissue Embedding, Journal Article",
author = "Christian Coupp{\'e} and Svensson, {Ren{\'e} B} and Heinemeier, {Katja M} and Thomsen, {Emilie W{\o}jdemann} and Bayer, {Monika Lucia} and Lise Christensen and Michael Kj{\ae}r and Magnusson, {S Peter} and Peter Schjerling",
year = "2017",
month = "1",
doi = "10.1007/s00418-016-1482-z",
language = "English",
volume = "147",
pages = "97--102",
journal = "Histochemistry and Cell Biology",
issn = "0948-6143",
publisher = "Springer",
number = "1",

}

RIS

TY - JOUR

T1 - Quantification of cell density in rat Achilles tendon

T2 - development and application of a new method

AU - Couppé, Christian

AU - Svensson, René B

AU - Heinemeier, Katja M

AU - Thomsen, Emilie Wøjdemann

AU - Bayer, Monika Lucia

AU - Christensen, Lise

AU - Kjær, Michael

AU - Magnusson, S Peter

AU - Schjerling, Peter

PY - 2017/1

Y1 - 2017/1

N2 - Increased tendon cell nuclei density (TCND) has been proposed to induce tendon mechanical adaptations. However, it is unknown whether TCND is increased in tendon tissue after mechanical loading and whether such an increase can be quantified in a reliable manner. The aim of this study was to develop a reliable method for quantification of TCND and to investigate potential changes in TCND in rat Achilles tendons in response to 12 weeks of running. Eight adult male Sprague-Dawley rats ran (RUN) on a treadmill with 10° incline, 1 h/day, 5 days/wk (17-20 m/min) for 12 weeks (which improved tendon mechanical properties) and were compared with 11 control rats (SED). Tissue-Tek-embedded cryosections (10 µm) from the mid region of the Achilles tendon were cut longitudinally on a cryostat. Sections were stained with alcian blue and picrosirius red. One blinded investigator counted the number of tendon cell nuclei 2-3 times in three separate regions of the mid longitudinal tendon sections with fields of 390 μm × 280 μm. Unpaired t tests were used for the statistical analysis (mean ± SE). Typical Error % for replicate counts was 5.5 and 14 % coefficient of variation for the three regions. There was no difference in TCND between running rats versus control rats (nuclei per image (≈105 μm2): RUN, 152 ± 9; SED, 146 ± 8, p = 0.642). This new method provided reproducible quantification of TCND. There was no difference in TCND despite improvements in tendon mechanics, which suggests that cell number is not a major cause for altered tendon mechanical properties with loading.

AB - Increased tendon cell nuclei density (TCND) has been proposed to induce tendon mechanical adaptations. However, it is unknown whether TCND is increased in tendon tissue after mechanical loading and whether such an increase can be quantified in a reliable manner. The aim of this study was to develop a reliable method for quantification of TCND and to investigate potential changes in TCND in rat Achilles tendons in response to 12 weeks of running. Eight adult male Sprague-Dawley rats ran (RUN) on a treadmill with 10° incline, 1 h/day, 5 days/wk (17-20 m/min) for 12 weeks (which improved tendon mechanical properties) and were compared with 11 control rats (SED). Tissue-Tek-embedded cryosections (10 µm) from the mid region of the Achilles tendon were cut longitudinally on a cryostat. Sections were stained with alcian blue and picrosirius red. One blinded investigator counted the number of tendon cell nuclei 2-3 times in three separate regions of the mid longitudinal tendon sections with fields of 390 μm × 280 μm. Unpaired t tests were used for the statistical analysis (mean ± SE). Typical Error % for replicate counts was 5.5 and 14 % coefficient of variation for the three regions. There was no difference in TCND between running rats versus control rats (nuclei per image (≈105 μm2): RUN, 152 ± 9; SED, 146 ± 8, p = 0.642). This new method provided reproducible quantification of TCND. There was no difference in TCND despite improvements in tendon mechanics, which suggests that cell number is not a major cause for altered tendon mechanical properties with loading.

KW - Achilles Tendon

KW - Animals

KW - Cell Count

KW - Cell Nucleus

KW - Male

KW - Rats

KW - Rats, Sprague-Dawley

KW - Tissue Embedding

KW - Journal Article

U2 - 10.1007/s00418-016-1482-z

DO - 10.1007/s00418-016-1482-z

M3 - Journal article

VL - 147

SP - 97

EP - 102

JO - Histochemistry and Cell Biology

JF - Histochemistry and Cell Biology

SN - 0948-6143

IS - 1

ER -

ID: 52334008