Histological composition and progression of carotid plaque
© Baroncini et al; licensee BioMed Central Ltd. 2007
Received: 06 December 2006
Accepted: 26 February 2007
Published: 26 February 2007
To analyse histological composition and progression of carotid plaque.
Thirty-one patients (22 males, mean age 68.03 ± 7.3 years) admitted for carotid endarterectomy for extracranial high-grade internal carotid artery stenosis (≥ 70% luminal narrowing) were enrolled. The patients were divided into 2 groups according to symptomatology (group I, 17 symptomatic patients; and group II, 14 asymptomatic patients). A histological analysis and inflammatory cell quantification of each excised carotid plaque was made. Nine carotid arteries were removed from human cadavers that were not preselected for carotid artery disease. These specimens were used as a control tissue without any macroscopic signs of atherosclerotic plaques.
Fifty eight percent of all carotid plaques were classified as complex plaque with possible surface defect, hemorrhage or thrombus. The inflammatory cells concentration did not differ between the two groups. All specimens from human cadavers were classified as preatheroma with extracellular lipid pools.
Asymptomatic and symptomatic patients could have the same histological components on their carotid plaques. Fibrotic and calcific plaques could become vulnerable as complex plaques with surface defect, hemorrhage and thrombus could remain silent. Asymptomatic carotid stenosis should be followed close with no invasive diagnostic methods and clinical evaluation.
In 1995, a report from the Committee on Vascular Lesions of the Council on Atherosclerosis, American Heart Association (AHA), had described the characteristic components and pathogenic mechanisms of the various advanced atherosclerotic lesions . This report provides a classification of human atherosclerotic lesions based on their histological composition and structure and reflects the temporal natural history of disease. The lesions were classified by Roman numerals that indicate the usual sequence of lesion progression, grading from type I (initial lesions) to type VIII (fibrotic plaque). Varying proportions of different components (connective tissue extracellular matrix; crystalline cholesterol, cholesteryl esters, phospholipids; and cells such as monocyte-derived macrophages, T lymphocytes, and smooth muscle cells) occur in different plaques, thus giving rise to a spectrum of lesions [2, 3]. Surface defects, hematoma, and thrombotic deposits futher damage, deform, and thicken, and accelerate conversion from clinically silent to overt disease . The present study was designed to characterize the progression and composition of carotid plaque, according with patient symptomatology, based on the American Heart Association (AHA) classification for human atherosclerotic lesions [1, 4]. The inflammatory cell quantification was based on estereology method.
Group I (n = 17) (symptomatic)
Group II (n = 14) (asymptomatic)
66.6 ± 6.7
67.6 ± 6.81
B. Procurement of tissue specimens and histological analysis
Carotid plaques were obtained immediately after endarterectomy. All surgeries were performed with standard surgical techniques, and with minimal manipulation of the specimen. No attempts were made to evaluate the presence and the degree of surface ulceration or thrombus. The plaque should be removed in bloc, without fragmentation or significant distortion. After removal, the section of plaque for histological analysis was placed in fresh 4% paraformaldehyde solution and partly decalcified overnight, in order to be sectioned subsequently. The samples were transected transversely at 3 to 4 mm, and embedded in paraffin. For the most of the specimens, five to six blocks were avaiable. Histological analysis was performed by an experience pathologist (SGR), based on American Heart Association classification for human atherosclerotic lesions [3, 4]. The inflammatory cell quantification was assessed by light microscopy with final magnification of 400 ×, using an ocular lens with a grid graticule. The observer selected a region in the plaque section with as many inflammatory cells as possible (hot spot). When no inflammatory area was clearly identified the selection was made in an aleatory way. Total area examined was 0.6 mm2 and each graticule grid corresponds to an area of 0,0625 mm2 with magnification of 400 ×. The inflammatory cells were counted in 10 different graticule area in each specimen. The human carotid arteries removed from cadavers had received the same histological treatment.
Continuous variables were expressed as mean ± SD. Statistical significance was indicated by a value of P < 0.05. The comparison of the histological parameters among the groups was done by non-parametric test of Kruskal-Wallis.
American Heart Association Classification for human atherosclerotic lesions according clinical groups.
Group I (n = 17) (symptomatic)
Group II (n = 14) (asymptomatic)
(minimum - maximum/specimen - median ± sd)
(22 ± 28)
(26 ± 29)
An asymptomatic patient with carotid artery stenosis is always a reason of concern for his personal physician. Current AHA guidelines recommended carotid endarterectomy (CEA) for asymptomatic patients, for stenosis 60% to 99%, if the risk of perioperative stroke or death is less than 3%. However, factors in addition to the degree of stenosis, such as the histological composition of the plaque, may be responsible for the determination of stroke risk. The mature atherosclerotic plaque is a complex structure suffering constantly of reparative process and its histological characterization is not easy. It was expected that types IV (atheroma with confluent extracellular lipid core), V (fibroatheroma) and VI (complex plaque with possible surface defect, hemorrhage or thrombus) of AHA classification should be the only ones found in group I (symptomatic patients), but we had classified two patients as having type VII (calcified plaque), and two patients as type VIII (fibrotic plaque). Also, in group II (asymptomatic patients) we found 11 patient as having plaques type VI. The fact that all surgical specimens in the present study had a high grade of complexity shows the challenge to try to separate vulnerable from stable carotid plaques. In the early stages of atherosclerosis the sequence is predictable, characteristics, and uniform, as we could evidence in the carotid arteries removed from human cadavers. However, lesions may subsequently progress in different morphogenetic sequences, resulting in several characteristic lesion types and clinical syndromes . It is well known that vulnerable plaques contain more total lipid and cholesterol, and less collagen and calcium as we could demonstrate in previous study [5–8]. However, the present study had evidenced that the atherosclerotic plaque is not a one-way progression from a "soft" to a "hard" plaque. When or why some these plaques will cross the line between stable to vulnerable? We know that fissures can occur in fibrotic and calcific plaques making then more vulnerable and the present study had evidenced that inflammatory cells are a constant part of atherosclerotic process, even in asymptomatic patients. The answer probably will be in the imunohistochemical analysis [9–14]. Most we have learned about vulnerable plaques is from imaging methods (ultrasound, magnetic resonance, and tomography) that wants to predict vascular events [15–29]. All these methods are able to identify precisely different proportions of fibrous tissue, lipid tissue and calcium as we had demonstrated in previous study . Some methods, mainly ultrasound tissue characterization can classify heterogeneous tissues based on second order statiscal parameters (entropy, homogeneity and energy) but they are failed in determine what this heterogeneous finds really means. Are they related with intrinsic reparative process inside the plaques? All these considerations suggest that asymptomatic carotid stenosis should be followed close with no invasive diagnostic methods and clinical evaluation.
First, the small number of patients was an important study limitation. This limitation will not be easily overcome, since the improvement of carotid artery stenting techniques will make histological analysis of carotid plaques an infrequent procedure. Second, by necessity, the plaques were sectioned and only a small proportion of each plaque was examined microscopically, and it may well be that features were missed in some patients. Most large lesions vary in composition along their length. This may particularly apply to the classification of fibrotic and calcific plaques in group I, where probably different components were missed when a small number of individual sections were examined. Third, we considered in this study patients with lacunar infarctions (LI) as had symptomatic carotid plaques. According to Tejeda et al , although significant carotid stenosis was observed at lower levels in LI, its pathogenic value should be taken into account because, when detected on the symptomatic side, it is not only a marker of atheromatosis but also a process potentially linked to LI. And finally, we did not perform imunohistochemical analysis in the present study that would certainly differentiate more instable carotid plaques, with large macrophage infiltration.
Mature carotid plaques are complex structures and their histological classification is a real challenge. Asymptomatic and symptomatic patients could have the same histological components on their carotid plaques. Fibrotic and calcific plaques could become vulnerable as complex plaques with surface defect, hemorrhage and thrombus could remain silent. Asymptomatic carotid stenosis should be followed close with no invasive diagnostic methods and clinical evaluation.
- Stary HC, Chandler AB, Dinsmore RE, Fuster V, Glagov S, Insull W, Rosenfeld ME, Schwartz CJ, Wagner WD, Wissler RW: A definition of advanced types of atherosclerotic lesions and a histological classification of atherosclerosis. Arterioscler Thromb Vasc Biol 1995, 15: 1512-1531.View ArticlePubMedGoogle Scholar
- Fayad ZA, Fuster V: Clinical imaging of the high-risk or vulnerable atherosclerotic plaque. Circ Res 2001, 89: 305-16.View ArticlePubMedGoogle Scholar
- Crisby M, Nordin-Fredriksson G, Shah PK, Yano J, Zhu J, Nilsson J: Pravastatin treatment increases collagen content and decreases lipid content, inflammation, metalloproteinases, and cell death in human carotid plaques. Implications for plaque stabilization. Circulation 2001, 103: 926-933.View ArticlePubMedGoogle Scholar
- Cai JM, Hatsukami TS, Ferguson MS, Small R, Polissar NL, Yuan C: Classification of human carotid atherosclerotic lesions with in vivo multicontrast magnetic resonance imaging. Circulation 2002, 106: 1368-1373. 10.1161/01.CIR.0000028591.44554.F9View ArticlePubMedGoogle Scholar
- Baroncini LAV, Pazin Filho A, Murta Junior LO, Martins AR, Ramos SG, Cherri J, Piccinato CE: Ultrasonic tissue characteriztion of vulnerable carotid plaque: correlation between videodensitometric method and histological examination. Cardiovascular Ultrasound 2006, 4: 32. 10.1186/1476-7120-4-32PubMed CentralView ArticlePubMedGoogle Scholar
- Wilhjelm JE, Grønholdt MLM, Wiebe B, Jespersen SK, Hansen LK, Sillesen H: Quantitative analysis of ultrasound B-mode imagesof carotid atherosclerotic plaque: correlation with visual classification and histological examination. IEEE Trans Med Imag 1998, 17: 910-922. 10.1109/42.746624View ArticleGoogle Scholar
- Takiuchi S, Rakugi H, Honda K, Masuyama T, Hirata N, Ito H, Sugimoto K, Yanagitani Y, Moriguchi K, Okamura A, Higaki J, Ogihara T: Quantitative ultrasonic tissue characterization can identify high-risk atherosclerotic alteration in human carotid arteries. Circulation 2000, 102: 766-770.View ArticlePubMedGoogle Scholar
- Tejada J, Díez-Tejedor E, Hernández-Echebarría L, Balboa O: Does a relationship exist between carotid stenosis and lacunar infarction? Stroke 2003, 34: 1404-1411. 10.1161/01.STR.0000072520.53106.8CView ArticlePubMedGoogle Scholar
- Cipollone F, Prontera C, Pini B, Marini M, Fazia M, De Cesare D, Iezzi A, Ucchino S, Boccoli G, Saba V, Chiarelli F, Cuccurullo F, Mezzetti A: Overexpression of functionally coupledcyclooxygenase-2 and prostaglandin E synthase in symptomaticatherosclerotic plaques as a basis of prostaglandin E(2)-dependent plaque instability. Circulation 2001, 104: 921-27.View ArticlePubMedGoogle Scholar
- Loftus IM, Naylor AR, Goodall S, Crowther M, Jones L, Bell PRF, Thompson MM: Increased Matrix Metalloproteinase-9 activity in unstable carotid plaques. A potencial role in acute plaque disruption. Stroke 2000, 31: 40-47.View ArticlePubMedGoogle Scholar
- Jander S, Sitzer M, Schumann R, Schroeter M, Siebler M, Steinmetz H, Stoll G: Inflammation in high-grade carotid stenosis. A possible role for macrophages and T cells in plaque destabilization. Stroke 1998, 29: 1625-1630.View ArticlePubMedGoogle Scholar
- Rothwell PM, Villagra R, Gibson R, Donders RC, Warlow CP: Evidence of a chronic systemic cause of instability of atherosclerotic plaques. Lancet 355(9197):19-24. 2000 Jan 1 10.1016/S0140-6736(99)04470-0Google Scholar
- Stoll G, Bendszus M: Inflammation and atherosclerosis. Novel insights into plaque formation and destabilization. Stroke 2006, 37: 1923-1932. 10.1161/01.STR.0000226901.34927.10View ArticlePubMedGoogle Scholar
- Redgrave JNE, Lovett JK, Gallagher PJ, Rothwell PM: Histological assessment of 526 symptomatic carotid plaques in relation to the nature and timing of ischemic symptoms. The Oxford plaque study. Circulation 2006, 113: 2320-2328. 10.1161/CIRCULATIONAHA.105.589044View ArticlePubMedGoogle Scholar
- Nighoghossian N, Derex L, Douek P: The vulnerable carotid artery plaque. Current imaging methods and new perspectives. Stroke 2005, 36: 2764-2772. 10.1161/01.STR.0000190895.51934.43View ArticlePubMedGoogle Scholar
- Geroulakos G, Ramaswami G, Nicolaides A, James K, Labropoulos N, Belcaro G, Holloway M: Characterization of symptomatic and asymptomatic carotid plaques using high-resolution real-time ultrasonography. Br J Surg 1993, 80: 1274-1277.View ArticlePubMedGoogle Scholar
- Nandalur KR, Baskurt E, Hagspiel KD, Phillips CD, Kramer CM: Calcified carotid atherosclerotic plaque is associated lesswith ischemic symptoms than is noncalcified plaque on MDCT. AJR 2005, 184: 295-298.PubMed CentralView ArticlePubMedGoogle Scholar
- Mathiesen EB, Bonaa KH, Joakimsen O: Echolucent plaques are associated with high risk of ischemic cerebrovascular events in carotid stenosis: the tromso study. Circulation 2001, 103: 2171-2175.View ArticlePubMedGoogle Scholar
- Gronholdt MLM, Nordestgaard BG, Schroeder TV, Vorstrup S, Sillesen H: Ultrasonic echolucent carotid plaques predict future strokes. Circulatio 2001, 104: 68-73.View ArticleGoogle Scholar
- Tegos TJ, Stavropoulos P, Sabetai MM, Khodabakhsh P, Sassano A, Nicolaides AN: Determinants of carotid plaque instability: echoicity versus heterogeneity. Eur J Vasc Endovasc Surg 2001, 22: 22-30. 10.1053/ejvs.2001.1412View ArticlePubMedGoogle Scholar
- Sabetai MM, Tegos TJ, Nicolaides AN, El-Atrozy TS, Dhanjil S, Griffin M, Belcaro G, Geroulakos G: Hemispheric symptoms and carotid plaque echomorphology. J Vasc Surg 2000, 31: 39-49. 10.1016/S0741-5214(00)70066-8View ArticlePubMedGoogle Scholar
- Pedro LM, Pedro MM, Gonçalves I, Carneiro TF, Balsinha C, Fernandes e Fernandes R, Fernandes e Fernandes J: Computer -assisted carotid plaque analysis: characteristics of plaques associated withcerebrovascular symptoms and cerebral infarction. Eur J Vasc Endovasc Surg 2000, 19: 118-123. 10.1053/ejvs.1999.0952View ArticlePubMedGoogle Scholar
- Liapis CD, Kakisis JD, Kostakis AG: Carotid stenosis. Factors affecting symptomatology. Stroke 2001, 32: 2782-2786.View ArticlePubMedGoogle Scholar
- Tegos TJ, Sohail M, Sabetai MM, Robless P, Akbar N, Pare G, Stansby G, Nicolaides AN: Echomorphologic and histopathologiccharacteristics of unstable carotid plaques. Am J Neuroradiol 2000, 21: 1937-1944.PubMedGoogle Scholar
- Mazzone AM, Urbani MP, Picano E, Paterni M, Borgatti E, DeFabritiis A, Landini L: In vivo ultrasonic parametric imaging of carotid atherosclerotic plaque by videodensitometric technique. Angiology 1995, 46: 663-672.View ArticlePubMedGoogle Scholar
- Beletsky VY, Kelley RE, Fowler M, Phifer T: Ultrasound densitometric analysis of carotid plaque composition. Stroke 1996, 27: 2173-2177.View ArticlePubMedGoogle Scholar
- Aly S, Bishop CC: An objective characterization ofatherosclerotic lesion. An alternative method to identify unstable plaque. Stroke 2000, 31: 1921-1924.View ArticlePubMedGoogle Scholar
- Lal BK, Hobson RW II, Pappas PJ, Kubicka R, Hameed M, Chakhtura EY, Jamil Z, Padberg FT Jr, Haser PB, Duran WN: Pixel distribution analysis of B – mode ultrasound scan images predicts histologic features of atherosclerotic carotid plaques. J Vasc Surg 2002, 35: 1210-1217. 10.1067/mva.2002.122888View ArticlePubMedGoogle Scholar
- Sztajel R, Momjian S, Momjian-Mayor I, Murith N, Djebaili K, Boissar G, Comelli M, Pizolato G: Stratified gray-scale median analysis and color mapping of the carotid plaque. Correlation with endarterectomy specimen histology of 28 patients. Stroke 2005, 36: 742-745.Google Scholar
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