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The analytical profile associated to a beveled specimen then is established in comparison with Stoney's formulae.
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Each specimen then was thermocycled in distilled water (3000 cycles) before immersion in sodium fluorescein dye for 24 hours.
The specimen then was subjected to a three-point bending force across the weakened area until the cartilage and subchondral bone fractured.
The specimens then were compressively loaded to failure in a universal testing machine.
Specimens then were washed, dehydrated, and embedded in Eponate 12 resin.
Specimens then were embedded in fresh O.C.T. and frozen in an ethanol/dry ice bath.
The specimens then were decalcified in 10% formic acid over 28 days and sectioned using a microtome.
The specimens then were fixed in zinc-buffered formalin (Z-Fix; Anatech Ltd ,Battle Creek, MI) at 4°C overnight.
Eligible cases and controls were contacted for permission to obtain their BBD pathology records and biopsy specimens, and specimens then were obtained from hospital pathology departments when possible.
The osteochondral specimens then were placed in standard cell culture conditions with DMEM for 4 days at 37°C in the incubator with 5% CO2.
The "enriched" specimen can then be used for insemination or in vitro fertilization.
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