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Technical Brief

Effect of Cold Storage and Freezing on the Biomechanical Properties of Swine Growth Plate Explants

[+] Author and Article Information
Anne-Laure Ménard

Dept. of Mechanical Engineering,
École Polytechnique of Montreal,
P.O. Box 6079, Station “Centre-Ville,”
Montréal, Québec H3C 3A7, Canada
Sainte-Justine University Hospital Center,
3175 Côte-Ste-Catherine Road,
Montréal, Québec H3T 1C5, Canada
e-mail: anne-laure.menard@polymtl.ca

Candice Soulisse

Dept. of Mechanical Engineering,
École Polytechnique of Montreal,
P.O. Box 6079, Station “Centre-Ville,”
Montréal, Québec H3C 3A7, Canada
Sainte-Justine University Hospital Center,
3175 Côte-Ste-Catherine Road,
Montréal, Québec H3T 1C5, Canada
e-mail: candice.soulisse@mines-saint-etienne.org

Pascale Raymond

Dept. of Mechanical Engineering,
École Polytechnique of Montreal,
P.O. Box 6079, Station “Centre-Ville,”
Montréal, Québec H3C 3A7, Canada
Sainte-Justine University Hospital Center,
3175 Côte-Ste-Catherine Road,
Montréal, Québec H3T 1C5, Canada
e-mail: pascale.raymond@polymtl.ca

Irène Londono

Sainte-Justine University Hospital Center,
3175 Côte-Ste-Catherine Road,
Montréal, Québec H3T 1C5, Canada
e-mail: irene.londono@recherche-ste-justine.qc.ca

Isabelle Villemure

Dept. of Mechanical Engineering,
École Polytechnique of Montreal,
P.O. Box 6079, Station “Centre-Ville,”
Montréal, Québec H3C 3A7, Canada
Sainte-Justine University Hospital Center,
3175 Côte-Ste-Catherine Road,
Montréal, Québec H3T 1C5, Canada
e-mail: isabelle.villemure@polymtl.ca

1Corresponding author.

Contributed by the Bioengineering Division of ASME for publication in the JOURNAL OF BIOMECHANICAL ENGINEERING. Manuscript received August 20, 2013; final manuscript received December 6, 2013; accepted manuscript posted December 13, 2013; published online March 24, 2014. Assoc. Editor: Guy M. Genin.

J Biomech Eng 136(4), 044502 (Mar 24, 2014) (5 pages) Paper No: BIO-13-1380; doi: 10.1115/1.4026231 History: Received August 20, 2013; Revised December 06, 2013; Accepted December 13, 2013

Ex vivo biomechanical testing of growth plate samples provides essential information about its structural and physiological characteristics. Experimental limitations include the preservation of the samples since working with fresh tissues involves significant time and transportation costs. Little information is available on the storage of growth plate explants. The aim of this study was to determine storage conditions that could preserve growth plate biomechanical properties. Porcine ulnar growth plate explants (n = 5 per condition) were stored at either 4 °C for periods of 1, 2, 3, and 6 days or frozen at −20 °C with slow or rapid sample thawing. Samples were tested using stress relaxation tests under unconfined compression to assess five biomechanical parameters. The maximum compressive stress (σmax) and the equilibrium stress (σeq) were directly extracted from the experimental curves, while the fibril-network reinforced biphasic model was used to obtain the matrix modulus (Em), the fibril modulus (Ef), and the permeability (k). No significant changes were observed in σeq and Em in any of the tested storage conditions. Significant decreases and increases, respectively, were observed in σmax and k in the growth plate samples refrigerated for more than 48 h and in the frozen samples, when compared with the fresh samples. The fibril modulus Ef of all stored samples was significantly reduced compared to the fresh samples. These results indicate that the storage of growth plates in a humid chamber at 4 °C for a maximum of 48 h is the condition that minimizes the effects on the measured biomechanical parameters, with only Ef significantly reduced. Refrigerating growth plate explants for less than 48 h maintains their maximal stress, equilibrium stress, matrix modulus, and permeability. However, cold storage at 4 °C for more than 48 h and freezing storage at −20 °C significantly alter the biomechanical response of growth plate samples. Appropriate growth plate sample storage will be beneficial to save time and reduce transportation costs to pick up fresh samples.

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Figures

Grahic Jump Location
Fig. 1

Growth plate sample processing. (a) Dissection of swine ulnae showing the distal epiphyseal growth plate; inset: a closer view of the explant shows the trimmed surface at the top after removal of the metaphyseal bony portion used for fixing the explant on the specimen holder. (b) Position of the disk sample between the two smooth platens of the micromechanical compression system. For the tests, the samples were bathed in Hank's balanced salt solution.

Grahic Jump Location
Fig. 2

Experimental stress relaxation curves from unconfined compression tests on growth plate explants. Black lines represent the values obtained for fresh samples. Gray lines represent the cold or frozen stored samples. The continuous line represents the mean values and the discontinuous ones the confidence intervals at 95%. (a)–(d) Cold stored samples for 24 (R1), 48 (R2), and 72 (R3) h and 6 days (R6), respectively. (e) and (f) Frozen stored samples at −20 °C, thawed at 37 °C (F20R), and thawed at 4 °C (F20S), respectively.

Grahic Jump Location
Fig. 3

Histogram bars for the mechanical stresses (a) and (b) and mechanical parameters obtained from the fibril-network-reinforced biphasic model (c)–(e) evaluated for all storage conditions: (a) σmax = maximum compressive stress, (b) σeq = equilibrium stress, (c) matrix modulus Em, (d) collagen fibril modulus Ef, and (e) permeability k. Means ± confidence intervals, n = 5 for each group

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