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Dimethyl sulfoxide toxicity kinetics in intact articular cartilage

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Abstract

Osteochondral defects can degenerate into osteoarthritis and currently there are no good treatment alternatives available to most Orthopaedic surgeons. Osteochondral allografting can restore damaged joint surfaces but its clinical use is limited by poor access to high quality tissue. Vitrification of osteochondral tissue would allow the banking of this tissue but requires high concentrations of cryoprotective agents. This study was designed to ascertain dimethyl sulfoxide (DMSO) toxicity kinetics to chondrocytes in situ after exposure to DMSO at different temperatures recorded as a function of time. Porcine osteochondral dowels were exposed to 1, 3, 5, and 6M DMSO at 4, 22, and 37°C for 0.5 min to 120 min. Chondrocyte recovery was determined by membrane integrity (Syto 13 and ethidium bromide) and mitochondrial (WST-1) assays. Results demonstrated that cell recovery was concentration, temperature and time dependent. At higher concentrations and temperatures, significant cell loss occurred within minutes. A rate constant calculated for chondrocyte death was dependent on temperature. 1 M DMSO appeared relatively non-toxic. This experiment established a method to examine systematically toxicity parameters for chondrocytes in situ and this data can be used to tailor vitrification protocols by limiting exposure temperature and time or lowering DMSO concentrations below toxic levels recorded.

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Abbreviations

AC:

Articular cartilage

ACI:

Autologous chondrocyte implantation

CGM:

Chondrocyte growth medium

CPA:

Cryoprotective agent

DMSO:

Dimethyl sulfoxide

EB:

Ethidium bromide

FDA:

Fluorescene diacetate

M:

molar

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

PBS:

Phospate-buffered saline

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Acknowledgements

This research was supported by Canadian Institutes of Health Research. The authors thank Sturgeon Valley Pork for supplying the joint tissue. J. A. W. Elliott holds a Canada Research Chair in Interfacial Thermodynamics. There are no professional or financial affiliations involved in preparation of this manuscript.

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Authors and Affiliations

  1. Department of Chemical and Materials Engineering, University of Alberta, Edmonton, T6G 2G6, Canada

    Janet A. W. Elliott

  2. Department of Surgery, University of Alberta Hospital, 2D2.32 WMC, 8440–112St, Edmonton, Alberta, T6G 2B7, Canada

    Anoop Poovadan, Garson K. Law & Nadr M. Jomha

  3. Department of Lab Medicine and Pathology, University of Alberta, Edmonton, T6G 2R8, Canada

    Heidi Y. Elmoazzen & Locksley E. McGann

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  1. Heidi Y. Elmoazzen
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  2. Anoop Poovadan
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  3. Garson K. Law
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  4. Janet A. W. Elliott
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  6. Nadr M. Jomha
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Correspondence to Nadr M. Jomha.

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Elmoazzen, H.Y., Poovadan, A., Law, G.K. et al. Dimethyl sulfoxide toxicity kinetics in intact articular cartilage. Cell Tissue Banking 8, 125–133 (2007). https://doi.org/10.1007/s10561-006-9023-y

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  • DOI: https://doi.org/10.1007/s10561-006-9023-y

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