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Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons

Frozen bone-patellar tendon bone allografts are useful in anterior cruciate ligament reconstruction as the freezing procedure kills tissue cells, thereby reducing immunogenicity of the grafts. However, a small portion of cells in human femoral heads treated by standard bone-bank freezing procedures...

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Autores principales: Suto, Kaori, Urabe, Ken, Naruse, Kouji, Uchida, Kentaro, Matsuura, Terumasa, Mikuni-Takagaki, Yuko, Suto, Mitsutoshi, Nemoto, Noriko, Kamiya, Kentaro, Itoman, Moritoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3286509/
https://www.ncbi.nlm.nih.gov/pubmed/21116722
http://dx.doi.org/10.1007/s10561-010-9234-0
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author Suto, Kaori
Urabe, Ken
Naruse, Kouji
Uchida, Kentaro
Matsuura, Terumasa
Mikuni-Takagaki, Yuko
Suto, Mitsutoshi
Nemoto, Noriko
Kamiya, Kentaro
Itoman, Moritoshi
author_facet Suto, Kaori
Urabe, Ken
Naruse, Kouji
Uchida, Kentaro
Matsuura, Terumasa
Mikuni-Takagaki, Yuko
Suto, Mitsutoshi
Nemoto, Noriko
Kamiya, Kentaro
Itoman, Moritoshi
author_sort Suto, Kaori
collection PubMed
description Frozen bone-patellar tendon bone allografts are useful in anterior cruciate ligament reconstruction as the freezing procedure kills tissue cells, thereby reducing immunogenicity of the grafts. However, a small portion of cells in human femoral heads treated by standard bone-bank freezing procedures survive, thus limiting the effectiveness of allografts. Here, we characterized the survival rates and mechanisms of cells isolated from rat bones and tendons that were subjected to freeze–thaw treatments, and evaluated the influence of these treatments on the mechanical properties of tendons. After a single freeze–thaw cycle, most cells isolated from frozen bone appeared morphologically as osteocytes and expressed both osteoblast- and osteocyte-related genes. Transmission electron microscopic observation of frozen cells using freeze-substitution revealed that a small number of osteocytes maintained large nuclei with intact double membranes, indicating that these osteocytes in bone matrix were resistant to ice crystal formation. We found that tendon cells were completely killed by a single freeze–thaw cycle, whereas bone cells exhibited a relatively high survival rate, although survival was significantly reduced after three freeze–thaw cycles. In patella tendons, the ultimate stress, Young’s modulus, and strain at failure showed no significant differences between untreated tendons and those subjected to five freeze–thaw cycles. In conclusion, we identified that cells surviving after freeze–thaw treatment of rat bones were predominantly osteocytes. We propose that repeated freeze–thaw cycles could be applied for processing bone-tendon constructs prior to grafting as the treatment did not affect the mechanical property of tendons and drastically reduced surviving osteocytes, thereby potentially decreasing allograft immunogenecity.
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spelling pubmed-32865092012-03-08 Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons Suto, Kaori Urabe, Ken Naruse, Kouji Uchida, Kentaro Matsuura, Terumasa Mikuni-Takagaki, Yuko Suto, Mitsutoshi Nemoto, Noriko Kamiya, Kentaro Itoman, Moritoshi Cell Tissue Bank Original Paper Frozen bone-patellar tendon bone allografts are useful in anterior cruciate ligament reconstruction as the freezing procedure kills tissue cells, thereby reducing immunogenicity of the grafts. However, a small portion of cells in human femoral heads treated by standard bone-bank freezing procedures survive, thus limiting the effectiveness of allografts. Here, we characterized the survival rates and mechanisms of cells isolated from rat bones and tendons that were subjected to freeze–thaw treatments, and evaluated the influence of these treatments on the mechanical properties of tendons. After a single freeze–thaw cycle, most cells isolated from frozen bone appeared morphologically as osteocytes and expressed both osteoblast- and osteocyte-related genes. Transmission electron microscopic observation of frozen cells using freeze-substitution revealed that a small number of osteocytes maintained large nuclei with intact double membranes, indicating that these osteocytes in bone matrix were resistant to ice crystal formation. We found that tendon cells were completely killed by a single freeze–thaw cycle, whereas bone cells exhibited a relatively high survival rate, although survival was significantly reduced after three freeze–thaw cycles. In patella tendons, the ultimate stress, Young’s modulus, and strain at failure showed no significant differences between untreated tendons and those subjected to five freeze–thaw cycles. In conclusion, we identified that cells surviving after freeze–thaw treatment of rat bones were predominantly osteocytes. We propose that repeated freeze–thaw cycles could be applied for processing bone-tendon constructs prior to grafting as the treatment did not affect the mechanical property of tendons and drastically reduced surviving osteocytes, thereby potentially decreasing allograft immunogenecity. Springer Netherlands 2010-11-30 2012 /pmc/articles/PMC3286509/ /pubmed/21116722 http://dx.doi.org/10.1007/s10561-010-9234-0 Text en © The Author(s) 2010 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Original Paper
Suto, Kaori
Urabe, Ken
Naruse, Kouji
Uchida, Kentaro
Matsuura, Terumasa
Mikuni-Takagaki, Yuko
Suto, Mitsutoshi
Nemoto, Noriko
Kamiya, Kentaro
Itoman, Moritoshi
Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
title Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
title_full Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
title_fullStr Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
title_full_unstemmed Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
title_short Repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
title_sort repeated freeze–thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3286509/
https://www.ncbi.nlm.nih.gov/pubmed/21116722
http://dx.doi.org/10.1007/s10561-010-9234-0
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