The use of pimonidazole to characterise hypoxia in the internal environment of an in vivo tissue engineering chamber

Abstract

The distribution of hypoxic cells in an in vivo tissue engineering chamber was investigated up to 28 days post-implantation. Methods: Arteriovenous loops were constructed and placed into bi-valved polycarbonate chambers containing 2×106 rat fibroblasts in basement membrane gel (BM gel). Chambers were inserted subcutaneously in the groin of male rats and harvested at 3 (n=6), 7 (n=6), 14 (n=4) or 28 (n=4) days. Ninety minutes before harvest, pimonidazole (60 mg/kg) was injected intraperitoneally. Chamber tissue was removed, immersion fixed, paraffin embedded, sectioned and stained immunohistochemically using hypoxyprobe-1 Mab that detects reduced pimonidazole adducts forming in cells, where p..