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Heart and Vessels
Published in: Heart and Vessels 1/2020

01-01-2020 | Varicosis | Original Article

Flow cytometric characterization of the saphenous veins endothelial cells in patients with chronic venous disease and in patients undergoing bypass surgery: an exploratory study

Authors: Cláudia Torres, Rui Machado, Margarida Lima

Published in: Heart and Vessels | Issue 1/2020

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Abstract

Recent findings have suggested that the primary factors for development of chronic venous disease (CVD), which commonly manifests as varicose veins (VV), are due to structural and biochemical modifications of the vessel wall. The aim of this exploratory study was to characterize by flow cytometry the endothelial cells (EC) mechanically extracted from the varicose saphenous veins (VSV) segments of patients submitted to VV surgery, and to compare the expression of cell surface molecules in these EC with that observed in the EC from the graft SV (GSV) of patients undergoing bypass surgery. EC were isolated from distal- (varicose trunk) and from proximal- (nearly normal) VSV segments of 30 patients submitted to VV surgery, and from proximal GSV segments of 20 patients submitted to bypass surgery (control group), using a mechanical method, and their immunophenotype was characterized by flow cytometry. EC were identified as being CD45negCD146brightCD31bright, and analyzed for expression of activation-related (CD54, CD62E, CD106), procoagulant (CD142), and cell junction (CD31, CD146) molecules, and for the scavenger receptor, CD36. The EC harvested from the SV segments of CVD patients had lower expression of all the molecules evaluated, in comparison to controls; these differences were more evident for the EC isolated from the distal-VSV. The EC extracted from the proximal- and distal-VSV segments of the CVD patients also differ from each other, the first having lower levels of CD62E, CD106, CD142 and CD36. Groups did not match for gender and controls were heterogeneous concerning the underlying pathologies, which may have a confounding effect. Our study revealed that the EC isolated from varicose (distal) and nearly normal (proximal) VSV segments of the CVD patients differ phenotypically from each other, and from the EC of the control group. The VSV segments more affected by the CVD have the lowest expression of the studied markers. We hypothesize that CVD is associated with a decrease on the EC surface molecules, causing EC dysfunctionality. Further studies with a large number of gender-matched participants are needed, to confirm the results obtained in this exploratory study.
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Literature
1.
Bergan JJ, Schmid-Schonbein GW, Smith PD, Nicolaides AN, Boisseau MR, Eklof B (2006) Chronic venous disease. N Engl J Med 355:488–498PubMed
2.
Beebe-Dimmer JL, Pfeifer JR, Engle JS, Schottenfeld D (2005) The epidemiology of chronic venous insufficiency and varicose veins. Ann Epidemiol 15:175–184PubMed
3.
Eberhardt RT, Raffetto JD (2014) Chronic venous insufficiency. Circulation 130:333–346PubMed
4.
Oklu R, Habito R, Mayr M, Deipolyi AR, Albadawi H, Hesketh R, Walker TG, Linskey KR, Long CA, Wicky S, Stoughton J, Watkins MT (2012) Pathogenesis of varicose veins. J Vasc Interv Radiol 23:33–39 (quiz 40) PubMed
5.
Evans CJ, Fowkes FG, Ruckley CV, Lee AJ (1999) Prevalence of varicose veins and chronic venous insufficiency in men and women in the general population: Edinburgh Vein Study. J Epidemiol Community Health 53:149–153PubMedPubMedCentral
6.
Canonico S, Gallo C, Paolisso G, Pacifico F, Signoriello G, Sciaudone G, Ferrara N, Piegari V, Varricchio M, Rengo F (1998) Prevalence of varicose veins in an Italian elderly population. Angiology 49:129–135PubMed
7.
Kontosic I, Vukelic M, Drescik I, Mesaros-Kanjski E, Materljan E, Jonjic A (2000) Work conditions as risk factors for varicose veins of the lower extremities in certain professions of the working population of Rijeka. Acta Med Okayama 54:33–38PubMed
8.
Criqui MH, Jamosmos M, Fronek A, Denenberg JO, Langer RD, Bergan J, Golomb BA (2003) Chronic venous disease in an ethnically diverse population: the San Diego Population Study. Am J Epidemiol 158:448–456PubMedPubMedCentral
9.
Clark A, Harvey I, Fowkes FG (2010) Epidemiology and risk factors for varicose veins among older people: cross-sectional population study in the UK. Phlebology 25:236–240PubMed
10.
Lim CS, Davies AH (2009) Pathogenesis of primary varicose veins. Br J Surg 96:1231–1242PubMed
11.
Cooper DG, Hillman-Cooper CS, Barker SG, Hollingsworth SJ (2003) Primary varicose veins: the sapheno-femoral junction, distribution of varicosities and patterns of incompetence. Eur J Vasc Endovasc Surg 25:53–59PubMed
12.
Elsharawy MA, Naim MM, Abdelmaguid EM, Al-Mulhim AA (2007) Role of saphenous vein wall in the pathogenesis of primary varicose veins. Interact Cardiovasc Thorac Surg 6:219–224PubMed
13.
Gandhi RH, Irizarry E, Nackman GB, Halpern VJ, Mulcare RJ, Tilson MD (1993) Analysis of the connective tissue matrix and proteolytic activity of primary varicose veins. J Vasc Surg 18:814–820PubMed
14.
Clarke GH, Vasdekis SN, Hobbs JT, Nicolaides AN (1992) Venous wall function in the pathogenesis of varicose veins. Surgery 111:402–408PubMed
15.
Venturi M, Bonavina L, Annoni F, Colombo L, Butera C, Peracchia A, Mussini E (1996) Biochemical assay of collagen and elastin in the normal and varicose vein wall. J Surg Res 60:245–248PubMed
16.
Sansilvestri-Morel P, Rupin A, Badier-Commander C, Kern P, Fabiani JN, Verbeuren TJ, Vanhoutte PM (2001) Imbalance in the synthesis of collagen type I and collagen type III in smooth muscle cells derived from human varicose veins. J Vasc Res 38:560–568PubMed
17.
Sansilvestri-Morel P, Rupin A, Jaisson S, Fabiani JN, Verbeuren TJ, Vanhoutte PM (2002) Synthesis of collagen is dysregulated in cultured fibroblasts derived from skin of subjects with varicose veins as it is in venous smooth muscle cells. Circulation 106:479–483PubMed
18.
Travers JP, Brookes CE, Evans J, Baker DM, Kent C, Makin GS, Mayhew TM (1996) Assessment of wall structure and composition of varicose veins with reference to collagen, elastin and smooth muscle content. Eur J Vasc Endovasc Surg 11:230–237PubMed
19.
Wali MA, Dewan M, Eid RA (2003) Histopathological changes in the wall of varicose veins. Int Angiol 22:188–193PubMed
20.
Wali MA, Eid RA (2002) Intimal changes in varicose veins: an ultrastructural study. J Smooth Muscle Res 38:63–74PubMed
21.
Badier-Commander C, Couvelard A, Henin D, Verbeuren T, Michel JB, Jacob MP (2001) Smooth muscle cell modulation and cytokine overproduction in varicose veins. An in situ study. J Pathol 193:398–407PubMed
22.
Demirkiran MA, Koksoy C, Heper AO, Bengisun U (2014) Does extracellular matrix of the varicose vein wall change according to clinical stage? Ulus Cerrahi Derg 30:186–191PubMedPubMedCentral
23.
Michiels C, Bouaziz N, Remacle J (2002) Role of the endothelium and blood stasis in the appearance of varicose veins. Int Angiol 21:1–8PubMed
24.
Saharay M, Shields DA, Georgiannos SN, Porter JB, Scurr JH, Coleridge Smith PD (1998) Endothelial activation in patients with chronic venous disease. Eur J Vasc Endovasc Surg 15:342–349PubMed
25.
Pocock ES, Alsaigh T, Mazor R, Schmid-Schonbein GW (2014) Cellular and molecular basis of Venous insufficiency. Vasc Cell 6:24PubMedPubMedCentral
26.
Poredos P, Jezovnik MK (2018) Endothelial dysfunction and venous thrombosis. Angiology 69(7):564–567PubMed
27.
Baggen VJ, Chung K, Koole K, Sarneel MH, Rutten FH, Hajer GR (2015) Association of varicosities and concomitant deep venous thrombosis in patients with superficial venous thrombosis, a systematic review. Eur J Gen Pract 21:70–76PubMed
28.
29.
Khaleel MS, Dorheim TA, Duryee MJ, Durbin HE Jr, Bussey WD, Garvin RP, Klassen LW, Thiele GM, Anderson DR (2012) High-pressure distention of the saphenous vein during preparation results in increased markers of inflammation: a potential mechanism for graft failure. Ann Thorac Surg 93:552–558PubMed
30.
Anderson CR, Hastings NE, Blackman BR, Price RJ (2008) Capillary sprout endothelial cells exhibit a CD36 low phenotype: regulation by shear stress and vascular endothelial growth factor-induced mechanism for attenuating anti-proliferative thrombospondin-1 signaling. Am J Pathol 173:1220–1228PubMedPubMedCentral
31.
Schuller-Petrovic S, Siedler S, Kern T, Meinhart J, Schmidt K, Brunner F (1997) Imbalance between the endothelial cell-derived contracting factors prostacyclin and angiotensin II and nitric oxide/cyclic GMP in human primary varicosis. Br J Pharmacol 122:772–778PubMedPubMedCentral
32.
Agu O, Hamilton G, Baker DM, Dashwood MR (2002) Endothelin receptors in the aetiology and pathophysiology of varicose veins. Eur J Vasc Endovasc Surg 23:165–171PubMed
33.
Bakke AC (2000) Clinical applications of flow cytometry. Lab Med 31:97–104
34.
Sack U, Tárnok A, Rothe G (2009) Cellular diagnostics: basic principles methods and clinical applications of flow cytometry. KARGER, Basel, pp 53–88
35.
Torres C, Machado R, Lima M (2016) Mechanical isolation of human endothelial cells from large vessels for flow cytometry immunophenotyping. J Tissue Sci Eng 7:1–6
36.
Bahr C (2007) CVI and PAD: a review of venous and arterial disease. JAAPA 20:20–25PubMed
37.
Orshal JM, Khalil RA (2004) Gender, sex hormones, and vascular tone. Am J Physiol Regul Integr Comp Physiol 286:R233–249PubMed
38.
Mudrovcic N, Arefin S, Van Craenenbroeck AH, Kublickiene K (2017) Endothelial maintenance in health and disease: Importance of sex differences. Pharmacol Res 119:48–60PubMed
39.
Tisato V, Zauli G, Voltan R, Gianesini S, di Iasio MG, Volpi I, Fiorentini G, Zamboni P, Secchiero P (2012) Endothelial cells obtained from patients affected by chronic venous disease exhibit a pro-inflammatory phenotype. PLoS ONE 7:e39543PubMedPubMedCentral
40.
Bachetti T, Morbidelli L (2000) Endothelial cells in culture: a model for studying vascular functions. Pharmacol Res 42:9–19PubMed
41.
Grange C, Letourneau J, Forget MA, Godin-Ethier J, Martin J, Liberman M, Latour M, Widmer H, Lattouf JB, Piccirillo CA, Cailhier JF, Lapointe R (2011) Phenotypic characterization and functional analysis of human tumor immune infiltration after mechanical and enzymatic disaggregation. J Immunol Methods 372:119–126PubMed
42.
Van Damme N, Baeten D, De Vos M, Demetter P, Elewaut D, Mielants H, Verbruggen G, Cuvelier C, Veys EM, De Keyser F (2000) Chemical agents and enzymes used for the extraction of gut lymphocytes influence flow cytometric detection of T cell surface markers. J Immunol Methods 236:27–35
43.
Manconi F, Markham R, Fraser IS (2000) Culturing endothelial cells of microvascular origin. Methods Cell Sci 22:89–99PubMed
44.
Jimenez N, Krouwer VJ, Post JA (2013) A new, rapid and reproducible method to obtain high quality endothelium in vitro. Cytotechnology 65:1–14PubMed
45.
Marin V, Kaplanski G, Gres S, Farnarier C, Bongrand P (2001) Endothelial cell culture: protocol to obtain and cultivate human umbilical endothelial cells. J Immunol Methods 254:183–190PubMed
46.
Fritschy JM (2008) Is my antibody-staining specific? How to deal with pitfalls of immunohistochemistry. Eur J Neurosci 28:2365–2370PubMed
47.
Birdina J, Pilmane M, Ligers A (2017) The morphofunctional changes in the wall of varicose veins. Ann Vasc Surg 42:274–284PubMed
48.
Takase S, Bergan JJ, Schmid-Schonbein G (2000) Expression of adhesion molecules and cytokines on saphenous veins in chronic venous insufficiency. Ann Vasc Surg 14:427–435PubMed
49.
Ghaderian SM, Lindsey NJ, Graham AM, Homer-Vanniasinkam S, Akbarzadeh Najar R (2010) Pathogenic mechanisms in varicose vein disease: the role of hypoxia and inflammation. Pathology 42:446–453PubMed
50.
Surendran S, Ramegowda SK, Suresh A, Binil Raj SS, Lakkappa RK, Kamalapurkar G, Radhakrishnan N, Kartha CC (2016) Arterialization and anomalous vein wall remodeling in varicose veins is associated with upregulated FoxC2-Dll4 pathway. Lab Invest 96:399–408PubMed
51.
52.
Jacobs BN, Andraska EA, Obi AT, Wakefield TW (2017) Pathophysiology of varicose veins. J Vasc Surg Venous Lymphat Disord 5:460–467PubMed
53.
Saharay M, Shields DA, Porter JB, Scurr JH, Coleridge Smith PD (1997) Leukocyte activity in the microcirculation of the leg in patients with chronic venous disease. J Vasc Surg 26:265–273PubMed
54.
Michiels C, Arnould T, Thibaut-Vercruyssen R, Bouaziz N, Janssens D, Remacle J (1997) Perfused human saphenous veins for the study of the origin of varicose veins: role of the endothelium and of hypoxia. Int Angiol 16:134–141PubMed
55.
Takase S, Schmid-Schonbein G, Bergan JJ (1999) Leukocyte activation in patients with venous insufficiency. J Vasc Surg 30:148–156PubMed
56.
Ferrero E, Ferrero ME, Pardi R, Zocchi MR (1995) The platelet endothelial cell adhesion molecule-1 (PECAM1) contributes to endothelial barrier function. FEBS Lett 374:323–326PubMed
57.
Ren Q, Ren L, Ren C, Liu X, Dong C, Zhang X (2015) Platelet endothelial cell adhesion molecule-1 (PECAM1) plays a critical role in the maintenance of human vascular endothelial barrier function. Cell Biochem Funct 33:560–565PubMed
58.
Bardin N, Anfosso F, Masse JM, Cramer E, Sabatier F, Le Bivic A, Sampol J, Dignat-George F (2001) Identification of CD146 as a component of the endothelial junction involved in the control of cell-cell cohesion. Blood 98:3677–3684PubMed
59.
Jacob MP, Cazaubon M, Scemama A, Prie D, Blanchet F, Guillin MC, Michel JB (2002) Plasma matrix metalloproteinase-9 as a marker of blood stasis in varicose veins. Circulation 106:535–538PubMed
60.
Sansilvestri-Morel P, Fioretti F, Rupin A, Senni K, Fabiani JN, Godeau G, Verbeuren TJ (2007) Comparison of extracellular matrix in skin and saphenous veins from patients with varicose veins: does the skin reflect venous matrix changes? Clin Sci (Lond) 112:229–239
61.
Mikula-Pietrasik J, Uruski P, Aniukiewicz K, Sosinska P, Krasinski Z, Tykarski A, Ksiazek K (2016) Serum from varicose patients induces senescence-related dysfunction of vascular endothelium generating local and systemic proinflammatory conditions. Oxid Med Cell Longev 2016:2069290PubMedPubMedCentral
62.
Budzyn M, Iskra M, Turkiewicz W, Krasinski Z, Gryszczynska B, Kasprzak MP (2018) Plasma concentration of selected biochemical markers of endothelial dysfunction in women with various severity of chronic venous insufficiency (CVI)-A pilot study. PLoS ONE 13:e0191902PubMedPubMedCentral
63.
Poredos P, Spirkoska A, Rucigaj T, Fareed J, Jezovnik MK (2015) Do blood constituents in varicose veins differ from the systemic blood constituents? Eur J Vasc Endovasc Surg 50:250–256PubMed
64.
Haviarova Z, Janegova A, Janega P, Durdik S, Kovac P, Stvrtinova V, Mraz P (2011) Nitric oxide synthases in varicose vein wall. Bratisl Lek Listy 112:18–23PubMed
65.
Xu Y, Bian X, Chu H, Zhao J, Wang T, Tang J, Guo W, Zhang S (2014) Effects of high hemodynamics upon the morphology of the walls of the great saphenous vein and splenic vein. Int Angiol 33:292–298PubMed
66.
Monici M (2005) Cell and tissue autofluorescence research and diagnostic applications. Biotechnol Annu Rev 11:227–256PubMed
67.
Chien S (2007) Mechanotransduction and endothelial cell homeostasis: the wisdom of the cell. Am J Physiol Heart Circ Physiol 292:H1209–1224PubMed
68.
Chiu JJ, Chien S (2011) Effects of disturbed flow on vascular endothelium: pathophysiological basis and clinical perspectives. Physiol Rev 91:327–387PubMed
69.
Yongbo X, Wei H, Lei W, Jianhua Z, Tao W, Jinyuan T, Kun L, Haibo C (2016) Changes in levels of apoptosis in the walls of different segments of great saphenous varicose veins. Phlebology 31:632–639PubMed
70.
Kun L, Ying L, Lei W, Jianhua Z, Yongbo X, Tao W, Jinyuan T, Haibo C (2016) Dysregulated apoptosis of the venous wall in chronic venous disease and portal hypertension. Phlebology 31:729–736PubMed
71.
Ascher E, Jacob T, Hingorani A, Tsemekhin B, Gunduz Y (2001) Expression of molecular mediators of apoptosis and their role in the pathogenesis of lower-extremity varicose veins. J Vasc Surg 33:1080–1086PubMed
72.
Surendran S, Girijamma A, Nair R, Ramegowda KS, Nair DH, Thulaseedharan JV, Lakkappa RB, Kamalapurkar G, Kartha CC (2014) Forkhead box C2 promoter variant c.-512C%3eT is associated with increased susceptibility to chronic venous diseases. PLoS One 9:e90682PubMedPubMedCentral
73.
Le Flem L, Mennen L, Aubry ML, Aiach M, Scarabin PY, Emmerich J, Alhenc-Gelas M (2001) Thrombomodulin promoter mutations, venous thrombosis, and varicose veins. Arterioscler Thromb Vasc Biol 21:445–451PubMed
Metadata
Title
Flow cytometric characterization of the saphenous veins endothelial cells in patients with chronic venous disease and in patients undergoing bypass surgery: an exploratory study
Authors
Cláudia Torres
Rui Machado
Margarida Lima
Publication date
01-01-2020
Publisher
Springer Japan
Published in
Heart and Vessels / Issue 1/2020
Print ISSN: 0910-8327
Electronic ISSN: 1615-2573
DOI
https://doi.org/10.1007/s00380-019-01451-9

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