Evaluation of tissue trauma and healing on the basis of tumour necrosis factor-alpha, interleukin-6 and C-reactive protein in peripheral blood during and after pyometra in bitches

Abstract

In this study, inflammation in the blood of bitches with pyometra (PG) was compared before and 15 days after ovariohysterectomy (OVH). The results were compared with those for control dioestrus bitches (CG) to reveal the evidence of inflammation in blood after a routine surgery. Inflammation was tracked by tumour necrosis factor-alpha (TNF-alpha), interleukine-6 (IL-6) and C-reactive protein (CRP), using immunocytochemistry (ICC) and immunofluorescence capture of blood cells in cell blocks. ICC is performed mainly during routine cytological examinations, whereas the use of cell blocks in blood examination is uncommon. Insofar as we know, this is the first study using cell block techniques on canine blood samples. Three commercially available antibodies against TNF-alpha, IL-6 and CRP, forming two panels, were evaluated. A standard streptoavidin-biotin complex technique was used for ICC. TNF-alpha and IL-6 labelling was scored for colour and intensity, and CRP for immunofluorescence capture. TNF-alpha and IL-6 colour and intensity scores differed significantly between the PG and CG groups, and were higher in PG before OVH (P < 0.01, P < 0.01, P < 0.001, P < 0.01, respectively). IL-6 intensity was significantly greater in PG 15 days after OVH (P < 0.05). CRP capture in PG was strong before OVH and high in both groups 15 days later. Low-dose anti-inflammatory agents or an anti-cytokine therapy may be useful in pyometra treatment in the future because these treatments may offer protection from systemic inflammatory response syndrome before and after OVH.