Dimethyl sulfoxide toxicity kinetics in intact articular cartilage.
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 degrees 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.
Elmoazzen
Heidi Y
HY
Department of Lab Medicine and Pathology, University of Alberta, Edmonton T6G 2R8, Canada.
Poovadan
Anoop
A
Law
Garson K
GK
Elliott
Janet A W
JA
McGann
Locksley E
LE
Jomha
Nadr M
NM
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects*; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects; D004121:Dimethyl Sulfoxide / Q000633:toxicity*; D007700:Kinetics; D013552:Swine; D013696:Temperature; D013997:Time Factors
D004121:Dimethyl Sulfoxide