A B S T R A C T This study was designed to investigate the mechanisms involved in fibromusculoelastic lesion formation produced by selective de-endothelialization by the intra-arterial balloon catheter technique in thrombocytopenic rabbits. Thrombocytopenia was induced and maintained for up to 31 days by daily injections of highly specific sheep anti-rabbit platelet sera (APS). Evidence for re-endothelialization was obtained by i.v. Evans blue dye 30 min before sacrifice. Rabbits received daily injections of APS, which reduced the mean platelet count to 5,600/cm3; control animals received identically treated normal sheep sera on the same schedule, and had mean daily platelet counts of 363,000/cm3. Evaluation of intimal thickness was assessed by counting cell layers in semithin sections. Intimal thickening in aortae from rabbits treated with APS was strikingly suppressed, in contrast to those from normal sheep sera-treated animals which showed a mean intimal thickness of 18 cell layers within 28 days often after de-endothelialization. Re-endothelialization was not affected by APS treatment. These results indicate that the proliferation of smooth muscle cells which is characteristic ofarteriosclerotic lesions is dramatically inhibited by reduction of platelets.

Arterial glycosaminoglycans are considered to be important in atherogenesis due to their ability to trap lipid inside the vessel wall and to influence cellular migration and proliferation. Atherosclerotic lesions have displayed an altered glycosaminoglycan content and distribution. Diabetes is a recognized risk factor for atherosclerosis, but no information is available on the arterial glycosaminoglycans in human diabetes. We examined glycosaminoglycans in normal and atherosclerotic intima of non-diabetic and Type 2 (non-insulin-dependent) diabetic patients. Intima was stripped from autopsy samples of thoracic aortas; normal and plaque areas were separated. Glycosaminoglycans were isolated by delipidation, proteolytic digestion, and precipitation and characterized by quantitation of total glycosaminoglycan and evaluation of glycosaminoglycan distribution by electrophoresis and densitometry. Results indicate a significant decrease in total glycosaminoglycan and significant changes in their distribution in atherosclerotic plaques: a relative decrease in heparan sulphate, a relative increase in dermatan sulphate and thus a decrease in the ratio of heparan sulphate to dermatan sulphate. A similar but less marked change in the ratio was found in normal intima of diabetic subjects, while in their plaques this change was more pronounced. This suggests that changes in arterial glycosaminoglycans (especially the ratio of heparan sulphate to dermatan sulphate) precede the development of lesions in diabetes and may be important in atherogenesis.

SummaryWe have previously shown that repeated or continuous intimal injury caused by an indwelling aortic catheter causes a variety of lesions in rabbits maintained on a diet unsupplemented by lipid. These include fatty streaks, lipid-free fibrous plaques and lipid-rich raised thromboatherosclerotic plaques. Whether lipid-rich raised lesions are a result of injury or co-existing thrombosis or both is not clear. The present experiment was designed to answer this question. Anti-platelet serum (APS) to washed sonicated rabbit platelets was raised in sheep. PE 60 polyethylene catheters were placed in the aortas of 35 rabbits by way of a femoral artery. The animals were randomly divided into 2 groups. The experimental group (17 rabbits) received an intravascular injection of 1.0 ml of APS followed 8 hours later by a subcutaneous injection of 0.5 ml. Thereafter, 0.5 ml APS was given subcutaneously each day for 13 additional days. The control group (18 rabbits) received no APS. Platelet counts were done prior to surgery, at 5 minutes following surgery, at 4 days, 8 days and just prior to killing. Extent of lesions was estimated by photographing the opened aortas, projecting the photographs on cardboard, cutting out the areas occupied by the different lesions and weighing the cardboard. The mean weight of raised lesions in the control group was 6 to 7 times greater than in the experimental groups. Statistical analysis of this difference based on Welsh‘s “t” test for unequal variances was highly significant (P<0.001). Platelet counts in the experimental group varied from 0 to 20,000 at 14 days. In animals with platelet counts ≤ 1,000 mm3 raised lesions were completely prevented. In a second experiment the effect of APS was compared with normal sheep serum (NSS). A similarly significant inhibition of raised lesions occurred in the APS group. The extent of lesions in the NSS control was similar to that in the No-APS group of the first experiment. These findings indicate that thrombosis is more important than injury in the development of lipid-rich raised lesions.

. Thromboatheromatous complications of umbilical arterial catheterization in the newborn period: clinicopathological study. Severe catheter-related thromboatheromatous lesions were found at necropsy in 33 of 56 infants who had umbilical arterial catheters passed during life. In infants dying within 8 days of insertion of the catheter, varying degrees of thrombosis of the aorta and its major branches were seen. With increasing thrombosis and aging of the thrombus, fatty deposits were seen first within the thrombus, and then in the intima and media. In addition there was evidence of proliferation of medial smooth muscle cells and of disruption of the medial architecture below the thrombus, characterized by the presence of abundant mucopolysaccharide. In infants who survived longer, varying degrees of organization of the thrombus could be traced, leading eventually to raised fibrous plaques with lipid and occasionally calcification. The lesions in the older infants were similar in many respects to experimental thromboatheromatous lesions produced in rabbits, and to some lesions of atheroma occurring spontaneously in humans.A wide variety of embolic phenomena were found, with features suggesting asynchrony of embolic episodes. The presence of thrombotic lesions could not be related to birthweight, Apgar scores at 1 and 5 minutes, age at catheterization, duration of catheterization, underlying disease process, age at death or the presence of hypothermia, acidosis, or anomalies in coagulation tests. There is a need for less hazardous methods of monitoring arterial oxygen tension.

We studied proteoglycan distribution In areas of spontaneously occurring high and low permeability by TEM examination of ruthenium red-stained sections of the aortic arch of normollpemlc and hyperllpemlc pigs. We noted granules of two sizes: those smaller than 20 nm contained heparan sulphate, and those from 20 to 50 nm In size contained chondroltln or dermatan sulphate. In the aortas of pigs fed a normal diet, there were significantly more granules of both types In low permeability areas than in areas permeable to Evans blue dye. This is consistent with the theory that glycosamlnoglycan provides a component for the control of aortic permeability. 4 It has been shown that permeable areas accumulate more cholesterol than contiguous, less permeable areas, when the animal studied has been fed a hypercholesterolemic diet.5 Such areas have generally been considered the sites where the initial stages of the atherosclerotic process develop. 6In lipid-fed animals, early atherogenic changes have been described in anatomical locations similar to those that are permeable to Evans blue in macroscopically normal vessels.16 " 8 More recently Gerrity et al. 9 have shown that, in the prelesion stages in swine fed an atherogenic diet, more monocytes accumulate on the endothelial surface of the areas per- meable to Evans blue dye than on the impermeable areas, and that lipid-filled monocytes were present in the intima of blue areas but not in the white areas. Subsequently, raised fatty lesions developed in the arch within the confines of the Evans blue dye uptake.10