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The effect of cryopreservation on canine menisci: a biochemical, morphologic, and biomechanical evaluation
S P Arnoczky1, C A McDevitt, M B Schmidt
1Laboratory of Comparative Orthopaedics, Hospital for Special Surgery, New York, NY 10021.
Abstract:
This study evaluated the effect of cryopreservation on the structural organization, biosynthetic activity, and material properties of canine menisci. The menisci were cryopreserved by incubating them in a 4% solution of dimethyl sulfoxide (DMSO) in physiologic media and freezing them to -100 degrees C using a controlled rate freezing system. The menisci were then stored for varying periods of time from zero to 12 weeks in liquid nitrogen (-196 degrees C). Following rapid thawing, changes in the histological appearance and biosynthetic activity of the menisci were evaluated as functions of storage time. In addition, the effects of the cryopreservation process on the tensile strength and modulus of the meniscal tissue were assessed. Although cryopreservation and short-term storage did not appear to affect the morphological appearance or biomechanical character of the menisci, biosynthetic activity, as determined by Na2S35SO4 incorporation, was diminished to less than 50% of normal control values immediately following cryopreservation and thawing. Autoradiographic examination of these tissues revealed that only approximately 10% of the meniscal cells were metabolically active, however, indicating that a marked increase in the metabolic activity of individual cells occurs following the freeze-thaw cycle. Total metabolic activity continued to decline with storage time.
Insights
Cryopreservation of canine menisci impacts biosynthetic activity, reducing it significantly post-thaw. While cell metabolism increases initially, overall tissue activity declines with extended storage.
Area of Science:
- Biomedical Engineering
- Tissue Engineering
- Veterinary Science
Background:
- Meniscal tears are common injuries in canines.
- Cryopreservation is a potential method for storing meniscal allografts.
- Understanding cryopreservation's effects on meniscal tissue is crucial for successful transplantation.
Purpose of the Study:
- To evaluate the impact of cryopreservation on canine menisci.
- To assess changes in structural, biosynthetic, and material properties.
- To determine the effects of storage duration on cryopreserved meniscal tissue.
Main Methods:
- Canine menisci were cryopreserved using dimethyl sulfoxide (DMSO) and controlled-rate freezing.
- Samples were stored in liquid nitrogen for 0-12 weeks.
- Histological appearance, Na2S35SO4 incorporation (biosynthetic activity), and tensile properties were assessed post-thaw.
Main Results:
- Cryopreservation and short-term storage did not alter meniscal morphology or biomechanical properties.
- Immediate post-thaw biosynthetic activity was reduced by over 50%.
- Only 10% of cells were metabolically active post-thaw, but showed increased individual cell activity; total activity decreased with storage time.
Conclusions:
- Cryopreservation affects the biosynthetic capacity of canine menisci.
- While structural and mechanical properties remain largely intact short-term, metabolic function is compromised.
- Further research is needed to optimize cryopreservation protocols to preserve meniscal cell viability and function.