July 18, 2023
American Journal Of Veterinary Research
Product: Botulinum Neurotoxin Type A from Clostridium botulinum
Cartilage explant culture and experimental design
A previously described in vitro model12 was used to simulate the effects of OA on articular cartilage. Under aseptic conditions, harvested cartilage was cut into 5 X 5 X 1 mm explants totaling 36 explants per joint (72 total explants). The wet weight of the explants was recorded. Explants were placed into individual wells of 12-well tissue culture plates with 2 mL of DMEM, supplemented as described above, for a 48-hour equilibration period at 5% CO2 and 37 °C. After the equilibration period, culture media was replaced with the treatment culture media of 1 of the 12 respective treatment groups (Table 1). Half of the explants were challenged with equine recombinant interleukin 1 (rEq IL-1; 0.1 ng/mL) to simulate osteoarthritis, and the other half remained unchallenged to simulate healthy joint conditions. Explants were exposed to 1 of 6 concentrations of research-grade BoNT-A (0, 1, 10, 50, 100, or 500 pg/mL; List Labs). Explants remained in culture for 96 hours with a media change at 48 hours.