Apoptosis in epithelial cells and its correlation with leukocyte accumulation in lamellar tissue from horses subjected to experimental sepsis-associated laminitis

Inflammation and apoptosis in the hoof lamellar interface both contribute to the early stages of sepsis-associated laminitis, but it is not clear whether apoptosis is occurring before the onset of inflammation or is being provoked by inflammation. Apoptosis and inflammation were therefore measured in lamellar tissues obtained at different time points throughout the early stages of experimentally induced laminitis. Apoptotic cells and leukocyte were enumerated in archived paraffin embedded lamellar tissue samples from previous experiments in which acute laminitis was induced using Black Walnut Extract (BWE) or starch (CHO). BWE-derived samples from 20 horses were allocated into four groups: Control (CON = 5); Early Time Point (ETP, 1.5 h after induction, n = 5); Developmental Time Point (DTP, 3-4 h after induction, n = 5); Obel Grade 1 (OG1, Onset of Lameness, n = 5). CHO-derived samples from 25 horses were allocated into four groups: CON (n = 8); DTP (10-12 h after induction, n = 6); OG 1 (n = 6); Obel 3 (OG3, lameness progression, n = 5). Apoptotic cells were enumerated using a horse validated TUNEL technique. Compared to controls, significant increases in apoptotic cell counts were not detected in lamellar epithelial cells during the developmental phase or at the onset of lameness during laminitis induction. A negative correlation between apoptosis and leukocyte infiltration was detected in the BWE model (P 0.05). In conclusion, apoptosis does not play an important role in the initial stages of sepsis-related laminitis.