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Chapter 6164Materials and Methods Following approval by the Medanex Clinic Ethics Committee (license number LA 1210576 - code of approval EC MxCl 2018-090), an in vivoexperimental animal study using fourteen ewes (Swifter crossbreed), was performed. A total of thirteen sheep were randomly divided into two groups. Both groups were implanted with a novel custom titanium 6-aluminum 4-vanadium (Ti6Al4V) TMJR system, of which six had a DLCcoated condyle. The other seven prosthesis had an uncoated condylar surface. One sheep functioned as a control group. In this case, the TMJ was surgically approached, yet no condylectomy or prosthesis implantation was performed. We refer to our previously published research for an extensive description of the surgery protocol and post-operative followup, and will only focus on the histological evaluation in this paper.(5,10) Sample processing and coloring288 days after implantation of the custom TMJR, all sheep were euthanized and decapitated. The skull was cut in half midsagitally. All the bodies and right half of the skulls were properly disposed of. The left half was further dissected by systematically removing the neurocranium, the anterior half of the mandible and maxilla, the upper half of the orbit and the orbital contents. The remaining tissue was then fixated for three months by immersion in 4% formaldehyde. Once properly fixated, all samples were rinsed for 3 days to remove the excess formalin. The peri-articular ‘neo-synovial’ tissues were then excised, taking care not to contain scar tissue from the implant surgery. During dissection of the neo-synovial tissues, several samples revealed an intracapsular brownish material, which appeared to be amorphous. This material was preserved and embedded in paraffin as well, to allow for further analysis. The ‘neo-synovial’ tissues were prepared, before staining, according to the following protocol; All specimen were put into sample cassettes and put into a 4% buffered Formalin solution. The samples were then washed out with running tap water to remove excess fixative from the tissues and prevent interaction of glutaraldehyde with the staining. The samples then were manually dehydrated in an ascending row of ethanol (30%, 50%, 60%, 70%) before further dehydration (90%, 96% ethanol and 100% Nikolas de Meurechy NW.indd 164 05-06-2024 10:14