Basement membrane pathology: a feature of acute equine laminitis

Abstract

Thirty-two dorsal, mid-hoof wall, lamellar sections from 8 Standardbred horses, humanely killed 48 h after the administration of an alimentary carbohydrate overload, were sectioned and examined by light microscopy. Sections were stained with the connective tissue and basement membrane stains periodic acid-Schiff (PAS), Azan and periodic acid silver methanamine (PASM) and with routine haematoxylin and eosin (H&E). Lesions of the epidermal lamellae, attributable to laminitis, were graded in order of increasing severity from Grade N (normal), Grade 1 (mild), Grade 2 (moderate) to Grade 3 (severe and extensive). The grading system was based principally on changes to lamellar basement membrane (BM) which were clearly visible when the connective tissue stains PAS and PASM were used. Earliest changes were rounding of the basal cell nuclei and elongation of secondary epidermal lamellae (SELs). Secondary epidermal lamellae tips were pointed instead of round and the basement membrane had separated from the lamellae. In early Grade 1 lesions, this was obvious at the tips of the SELs where the BM had lifted to form teat-shaped bubbles. The absence of BM at the tips of secondary dermal lamellae, along with varying amounts of connective tissue, was considered a progression in severity and classified as Grade 2. Eventually, even the primary epidermal and primary dermal lamellae separated from each other and the empty shells of isolated BM, in what was once the tip of the primary epidermal lamella, signified that a global separation of the epidermal and dermal lamellae had occurred (Grade 3 lesion). The histopathological grading system correlated well with the degree of lameness at the time of euthanasia, (r2 = 0.94) and apparently described the severity of laminitis accurately. Disintegration of the BM and failure of its attachment to the basal cells of the epidermis appears to be one of the earliest pathological events to occur in acute laminitis and could be the change that initiates the collapse of the lamellar architecture. Histopathological diagnoses of laminitis are strengthened when based on sections stained with at least PAS, in addition to routine H&E and should exhibit evidence of the BM pathology described here.