Changes in Adrenergic Nerves and Tissue Perfusion after Freezing Injury to the Ear Skin of Rabbits

Abstract

The aim of the present study was to investigate by a combined trypan blue and catecholamine fluorescence technique the sequential changes in cutaneous adrenergic nerves, the distribution of blood flow, and vascular permeability at the demarcation line that occurred after experimental cold injury. Nine New Zealand white rabbits weighing 4.0-4.4 kg had frostbite induced by pressing the bottom of a glass bottle 2 cm in diameter filled with liquid nitrogen against the shaved skin of the ear. All the rabbits were anaesthetised with ketamine hydrochloride (Ketamine®) and xylazine hydrochloride (Rompun®). Specimens were taken one and three days, and two weeks after frostbite. Control samples were taken from the opposite normal ear. Trypan blue was injected into a saphenous vein just before the specimens were taken. The specimens were always taken in the same way and selected so that the probable demarcation line of the frostbite ran across the middle. The glyoxylic acid-induced fluorescence method was used to show the adrenergic nerves. There were no adrenergic nerves around the vessels and no arteriovenous anastomoses in the central area of the injury after one day, but catecholamines had started to accumulate in the adrenergic nerve endings at the margins of the injured area. This accumulation was still more obvious three days after frostbite. Some fluorescent regenerating adrenergic nerves could already be seen at the probable demarcation line two weeks after frostbite. There was increased trypan blue fluorescence near the margin of the injured area after one day, indicating extravasation and vascular damage. This red fluorescence was more evident at the margins three days after frostbite, but it was no longer so intense after two weeks.

Key Words:

• histofluorescence
• trypan blue
• frostbite
• demarcation line