Microscopic observations on amputated and scarring lizard digits show an intense inflammatory reaction

Abstract

The microscopic details of the failure of digit regeneration in lizards are not known. The present study reports some histological, ultrastructural and 5BrdU-immunohistochemical observations on healing digits after amputation in the lizard Podarcis muralis. At 7-12 days post-amputation, the stump of digits forms a multilayered wound epidermis covering a loose connective tissue that is invaded by granulocytes, macrophages and lymphocytes. In addition to macrophages also electron-pale multinuclear giant cells are seen underneath or penetrating the wound epidermis while osteoclasts are present in the degrading bone of the severed phalanges. Granulocytes and macrophages invading the wound epidermis indicate the formation of an intra-epidermal immune barrier beneath the scab where numerous bacteria remain entrapped. Immunofluorescence for 5BrdU reveals that few proliferating cells are present in the wound epidermis and the underlying connective tissue at 12 and 32 days post-amputation. Outgrowths of less than 1mm stop growing and at 32 days they appear scaling. Most of connective cells give rise to fibrocytes and large irregular collagen bundles, as is typical for scar tissue. In conclusion, like for the amputated limb, the intense inflammatory reaction and scarring here described after digit loss appears associated with immune cells invasion.