The Quantitation of Hemorrhage in the Skin. Measurement of Hemorrhage in the Microcirculation in Inflammatory Lesions and Related Phenomena

Abstract

The acute inflammatory reaction has been extensively studied and several of its parameters quantitated. The most thorough measurements involved changes in blood flow, vascular permeability, and leukocyte infiltration. Although hemorrhage has often been observed in inflammatory reactions, to our knowledge, no satisfactory technique has been developed which is sensitive enough to quantitate it in cutaneous lesions. However, hemorrhage into soft tissues can be quantitated as shown by Schertzer and Ward ( 1 ) by homogenizing the tissue, and, following extraction, estimating the h e m o g l o b i n c o n c e n t r a t i o n s p e ctrophotometrically . A similar approach was attempted in our laboratory. Hemoglobin was extracted from hemorrhagic skin lesions by the method of Dodge et al.(2), but this method proved to be too insensitive in our system. A second approach was to use "Cr or '"In to label red blood cells (RBCs) in vitro followed by their reinfusion. These techniques proved feasible, but sensitivity of the assay was greatly improved when RBCs were labeled in virw with .' "e. In initial experiments the RBCs were labeled in the rabbits in which dermal lesions were induced. Subsequently the RBCs of a donor rabbit were labeled in vivo and then transfused into recipient rabbits. Hemorrhagic lesions were induced in the skin of recipient

Quantitation of Platelets in the Microcirculation. Measurement of Indium-111 in Microthrombi Induced in Rabbits by Inflammatory Lesions and Related Phenomena

“…However, one of the complications of an inflammatory reaction is thrombosis of the microcirculation. Hemorrhage is often associated with bacterial inflammatory lesions and with lesions such as the local Shwartzman reaction (19). Thrombi in the microcirculation may also be present in both these reactions.…”

“…It is now possible to study the kinetics of the acute inflammatory reaction (16), by quantitating the infiltration of neutrophils (17,18), the amount of hemorrhage (19), the increase in vascular permeability (20), and alternations in blood flow in the microcirculation (21). However, one of the complications of an inflammatory reaction is thrombosis of the microcirculation.…”

“…Two types of leisons were elicited. Inflammatory lesions were produced as described before (16). Briefly, the hair of the back of 2-2.5 kg New Zealand white rabbits was clipped and 6 x los Formalin-killed Escherichia coli (BAM strain, Medical Teaching Laboratories, University of Toronto, Toronto, Ontario, Canada) were injected intradermally (0.2 ml) at the following times prior to sacrifice: 24, 8, 6, 4, 3, 2, and 1 hr.…”

“…The fate of the platelets in the circulation was followed by taking blood samples at 5 and 50 min after the infusion of the platelet suspension. After killing the rabbits with an overdose of sodium pentobarbital, the skin of the back was removed, the visible blood was expressed manually from the large vessels (19), and the lesions were punched out with a cork borer (1.5 cm in diameter). The disks of skin were counted in the y-scaler.…”

Application of ⁵¹Cr-Labelled PMN Leukocytes in Quantitating PMN Kinetics in Systemic and Local Inflammatory Mediated Processes

“…A 20 pg/kg dose of a more potent endotoxin (E. coli 055: B5 Difco Laboratories, Detroit, Mich.; lot 682197) was injected intravenously into 4 rabbits in which 5,Cr-PMNs were circulating for 100 min and a 5th rabbit served as con trol. Each rabbit received 59Fe erythrocytes [Cybulsky and Moral, 1982;Kopaniak et al, 1980a] from a donor rabbit 5 min prior to sacrifice and a blood sample was taken im mediately before sacrifice. The control rabbit was sacrificed 100 min after injection of 5,Cr-PMN and the other rabbits 5, 15, 30 and 90 min later.…”