D4 structurix dw ete

Immediately after harvesting the biopsies for bacteriology, we placed the infected legs containing the titanium plate and surrounding soft tissue in 70% methanol. Contact radiographs were taken using a cabinet X-ray system (Model No. 43855A, Faxitron X-Ray Corp) and high-resolution technical films (D4 Structurix DW ETE, Agfa). After fixation, samples were subjected to a dehydration process through ascending concentrations of ethanol and transferred to xylene before being embedded in methyl methacrylate (Sigma-Aldrich). Once cured, two approximately 200-µm-thick longitudinal sections were made through the plate and femur using a Leica 1600 rotating saw microtome (Leica microsystems). These sections were fixed with cyanoacrylate onto Plexiglas slides and ground to a thickness of approximately 100 μm using a microgrinding system. One section per animal was stained with Giemsa eosin.

Sixteen animals were operated to provide sections for histopathological evaluation of infected and control animals (n = 4 per group, per time-point). Animals were sacrificed at 3 and 7 days to evaluate the acute phase of the infection. Immediately after euthanasia, the entire operated femur and surrounding musculature underwent immediate fixation with 70% (v/v) methanol. Contact radiographs were taken of the entire specimen using high resolution technical films (D4 Structurix DW ETE, Agfa, Belgium) and a cabinet X-ray system (Model No. 43855A, Faxitron X-Ray Corporation, USA). Following fixation, samples were dehydrated through an ascending series of ethanol, transferred to xylene, and finally infiltrated with and embedded in polymethylmethacrylate (PMMA) [25] . The polymerized blocks were cut using an annular blade saw (Leitz 1600 saw microtome, Leica AG, Switzerland) resulting in 1 longitudinal cut through the plate per femur. Finally, each section was glued with cyanoacrylate onto plexiglass slides, ground and polished down to approximately 100 μm using a micro-grinding system (Exact®, Germany) and stained with Giemsa/Eosin (GE).

Mediolateral and craniocaudal radiographic projections of the left tibia of each sheep were taken before each surgery, immediately after each surgery, then every other week and at euthanasia. Sheep were sedated for radiographs (0.04 mg/kg i.m. of detomidine; E. Graeub AG, Bern, Switzerland). Additionally, a high-resolution contact radiograph (full thickness) was taken post-mortem, using highresolution technical film (D4 Structurix DW ETE, Agfa ® , Belgium) and a cabinet X-ray system (Model No. 43855A, Faxitron X-Ray Corporation ® , USA) at 42 kV for 5 min, with a 0.5 mm aluminium filter.

Rabbit humeri, including implant and adjacent soft tissue, were fixed in 70% (v/v) methanol for a minimum of 2 weeks with fresh methanol changes weekly. Contact radiographs (full thickness) were taken using high resolution technical film (D4 Structurix DW ETE, Agfa, Belgium) and a cabinet X-ray system (Model No. 4385A, Faxitron X-ray Corporation, AZ, USA). After fixation, samples were dehydrated by an ascending series of ethanol and were transferred to xylene. Finally, they were infiltrated and embedded in methylmethacrylate (MMA). The polymerized MMA blocks were trimmed using a butcher saw (Bizerba FK 22, Bizerba AG, Switzerland) prior to cutting with an annular diamond saw (Leitz 1600 saw microtome, Leica AG, Switzerland). The samples were glued with cyanoacrylate onto Beracryl holders for sectioning. Two sections of each sample were selected, which were glued onto opaque Plexiglass Ò slides, ground and fine polished. Sections were stained with Giemsa-Eosin and histopathological analysis of the slides was performed using a transmission light microscope (BX40, Olympus, Switzerland). Histological findings were described, wherever possible, according to distribution (focal, multifocal, diffuse), morphological character and to severity by a veterinary pathologist.