Environmental Science and Pollution Research

, Volume 26, Issue 1, pp 991–999 | Cite as

Pure ultra-fine carbon particles do not exert pro-coagulation and inflammatory effects on microvascular endothelial cells

  • Hossein Dinmohammadi
  • Zahra Pirdel
  • Laleh Salarilak
  • Marc Hoylaerts
  • Reza Nejatbakhsh
  • Alireza Biglari
  • Marc Jacquemin
  • Tina Shahani
Short Research and Discussion Article


Pro-thrombotic and inflammatory changes play an important role in cardiovascular morbidity and mortality, resulting from short-term exposure to fine particulate air-pollution. Part of those effects has been attributed to the ultra-fine particles (UFPs) that pass through the lung and directly contact blood-exposed and circulating cells. Despite UFP-induced platelet activation, it is unclear whether the penetrated particles exert any direct effect on endothelial cells. While exposure levels are boosting as a result of world-wide increases in economic development and desertification, which create more air-polluted regions, as well as increase in demands for synthetic UFPs in medicine and various industries, further studies on the health effects of these particles are required. In this study, human pulmonary and cardiac microvascular endothelial cells (MECs) have been exposed to 0.1, 1, 10, and 100 μg/ml suspensions of either a natural (carbon black) or a synthetic (multi-walled carbon nano-tubes) type of UFPs, in vitro. As a result, no changes in the levels of coagulation factor VIII, Von Willebrand factor, Interleukin 8, and P-selectin measured in the cells’ supernatant were observed prior to and 6, 12, and 24 h after exposure. In parallel, the spatio-temporal effect of UFPs on cardiac MECs was evaluated by Transmission Electron Microscopy. Despite phagocytic uptake of pure UFPs observed on cellular sections of the treated cells, Weibel-Palade bodies remained intact in shape and similar in number when compared with the untreated cells. Our work shows that carbon itself is a non-toxic carrier for endothelial cells.


Human cardiac microvascular endothelial cells Human pulmonary microvascular endothelial cells FVIII VWF IL-8 P-selectin Weibel-Palade bodies 



Coagulation factor VIII


FVIII activity


Human cardiac microvascular endothelial cells


Human pulmonary microvascular endothelial cells


Interleukin 8


Limit of detection


Particulate matter


Phorbol-12-myristate 13-acetate


Ultra-fine particle


Von-Willebrand factor


Weibel-Palade body


Transmission electron microscopy



Special thanks to Mrs. Zahra Ramezani for her technical support with TEM at Biomedical Engineering and Medical Physics Department of Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Authors’ contributions

HD, MH, and TSH wrote the manuscript. HD, ZP, LS, and RN performed/analyzed the experiments. MH, AB, MJ, and TSH contributed to the study design and data analysis. All authors read and approved the final manuscript.

Funding information

This work is supported by Zanjan University of Medical Sciences (ZUMS) grant number A-12-776-2 and Iran’s National Institute for Medical Research Development (NIMAD) grant number 940200.

Compliance with ethical standards

Ethics approval

The study was conducted in accordance with the Helsinki Declaration and approved by Zanjan University of Medical Sciences Ethics Committee (ZUMS.REC.1394.132; 01-Sept 2015).

Consent for publication

Not applicable.

Availability of data and material

The data that support the findings of this study are available from the corresponding authors on reasonable request.

Competing interests

The authors declare that they have no conflict of interest.

Supplementary material

11356_2018_3783_MOESM1_ESM.pdf (62 kb)
Figure S1 Effect of CB and MWCNTs on endothelial cell viability assayed by MTT test. Cells were incubated with 10 and 100 μg/ml of CB or MWCNTs for 24 h. The values for cell viability represent the mean of three experiments shown as percentage to the untreated cells, indicated as controls. Error bars represent standard deviation (SD). The abbreviations are: CB (Carbon Black), MWCNTs (Multi-wall Carbon Nano-Tube). * P value <0.001. (PDF 62 kb)


  1. Abdalla S, Al-Marzouki F, Al-Ghamdi AA, Abdel-Daiem A (2015) Different technical applications of carbon nanotubes. Nanoscale Res Lett 10:358CrossRefGoogle Scholar
  2. Adams K, Greenbaum DS, Shaikh R, van Erp AM, Russell AG (2015) Particulate matter components, sources, and health: systematic approaches to testing effects. J Air Waste Manag Assoc 65:544–558. CrossRefGoogle Scholar
  3. Adams WJ, Zhang Y, Cloutier J, Kuchimanchi P, Newton G, Sehrawat S, Aird WC, Mayadas TN, Luscinskas FW, García-Cardeña G (2013) Functional vascular endothelium derived from human induced pluripotent stem cells. Stem Cell Rep 1:105–113CrossRefGoogle Scholar
  4. Brook RD, Rajagopalan S, Pope CA 3rd, Brook JR, Bhatnagar A, Diez-Roux AV, Holguin F, Hong Y, Luepker RV, Mittleman MA, Peters A, Siscovick D, Smith SC Jr, Whitsel L, Kaufman JD, American Heart Association Council on Epidemiology and Prevention, Council on the Kidney in Cardiovascular Disease, and Council on Nutrition, Physical Activity and Metabolism (2010) Particulate matter air pollution and cardiovascular disease: an update to the scientific statement from the American Heart Association. Circulation 121:2331–2378CrossRefGoogle Scholar
  5. Brutsaert DL, Meulemans AL, Sipido KR, Sys SU (1988) Effects of damaging the endocardial surface on the mechanical performance of isolated cardiac muscle. Circ Res 62:358–366CrossRefGoogle Scholar
  6. Devouassoux G, Saxon A, Metcalfe DD, Prussin C, Colomb MG, Brambilla C, Diaz-Sanchez D (2002) Chemical constituents of diesel exhaust particles induce IL-4 production and histamine release by human basophils. J Allergy Clin Immunol 109:847–853CrossRefGoogle Scholar
  7. Dominici F, Peng RD, Bell ML, Pham L, McDermott A, Zeger SL, Samet JM (2006) Fine particulate air pollution and hospital admission for cardiovascular and respiratory diseases. Jama 295:1127–1134CrossRefGoogle Scholar
  8. Emmerechts J, Hoylaerts M (2012) The effect of air pollution on haemostasis. Hämostaseologie 32:5–13CrossRefGoogle Scholar
  9. Franciosi S, Perry FK, Roston TM, Armstrong KR, Claydon VE, Sanatani S (2017) The role of the autonomic nervous system in arrhythmias and sudden cardiac death. Autonomic Neuroscience: Basic and Clinical 205:1–11CrossRefGoogle Scholar
  10. Ghio AJ, Devlin RB (2001) Inflammatory lung injury after bronchial instillation of air pollution particles. Am J Respir Crit Care Med 164:704–708. CrossRefGoogle Scholar
  11. Gurgueira SA, Lawrence J, Coull B, Murthy GG, Gonzalez-Flecha B (2002) Rapid increases in the steady-state concentration of reactive oxygen species in the lungs and heart after particulate air pollution inhalation. Environ Health Perspect 110:749–755CrossRefGoogle Scholar
  12. Gwinn MR, Vallyathan V (2006) Nanoparticles: health effects—pros and cons. Environ Health Perspect 114:1818–1825CrossRefGoogle Scholar
  13. Hadi HA, Carr CS, Al Suwaidi J (2005) Endothelial dysfunction: cardiovascular risk factors, therapy, and outcome Vascular health and risk management 1:183Google Scholar
  14. He R-W, Shirmohammadi F, Gerlofs-Nijland ME, Sioutas C, Cassee FR (2018) Pro-inflammatory responses to PM 0.25 from airport and urban traffic emissions. Sci Total Environ 640:997–1003CrossRefGoogle Scholar
  15. Hei, H (2013) Review panel on ultrafine particles. Understanding the health effects of ambient ultrafine particles HEI Perspectives 3Health Effects Institute, Boston, MA:122Google Scholar
  16. Hertel S, Viehmann A, Moebus S, Mann K, Bröcker-Preuss M, Möhlenkamp S, Nonnemacher M, Erbel R, Jakobs H, Memmesheimer M, Jöckel KH, Hoffmann B (2010) Influence of short-term exposure to ultrafine and fine particles on systemic inflammation. Eur J Epidemiol 25:581–592CrossRefGoogle Scholar
  17. Hoet PH, Brüske-Hohlfeld I, Salata OV (2004) Nanoparticles–known and unknown health risks. J Nanobiotechnol 2(1):12CrossRefGoogle Scholar
  18. Huang W, Wang G, Lu SE, Kipen H, Wang Y, Hu M, Lin W, Rich D, Ohman-Strickland P, Diehl SR, Zhu P, Tong J, Gong J, Zhu T, Zhang J (2012) Inflammatory and oxidative stress responses of healthy young adults to changes in air quality during the Beijing Olympics. Am J Respir Crit Care Med 186:1150–1159. CrossRefGoogle Scholar
  19. Jacquemin M, Neyrinck A, Hermanns MI, Lavend'homme R, Rega F, Saint-Remy JM, Peerlinck K, van Raemdonck D, Kirkpatrick CJ (2006) FVIII production by human lung microvascular endothelial cells. Blood 108:515–517CrossRefGoogle Scholar
  20. Jiang N, Dreher KL, Dye JA, Li Y, Richards JH, Martin LD, Adler KB (2000) Residual oil fly ash induces cytotoxicity and mucin secretion by guinea pig tracheal epithelial cells via an oxidant-mediated mechanism. Toxicol Appl Pharmacol 163:221–230CrossRefGoogle Scholar
  21. Karoly ED, Li Z, Dailey LA, Hyseni X, Huang Y-CT (2007) Up-regulation of tissue factor in human pulmonary artery endothelial cells after ultrafine particle exposure. Environ Health Perspect 115:535–540CrossRefGoogle Scholar
  22. Khandoga A, Stampfl A, Takenaka S, Schulz H, Radykewicz R, Kreyling W, Krombach F (2004) Ultrafine particles exert prothrombotic but not inflammatory effects on the hepatic microcirculation in healthy mice in vivo. Circulation 109:1320–1325CrossRefGoogle Scholar
  23. Kido T, Tamagawa E, Bai N, Suda K, Yang HHC, Li Y, Chiang G, Yatera K, Mukae H, Sin DD, van Eeden SF (2011) Particulate matter induces translocation of IL-6 from the lung to the systemic circulation. Am J Respir Cell Mol Biol 44:197–204CrossRefGoogle Scholar
  24. Kilinc E et al (2011) Factor XII activation is essential to sustain the procoagulant effects of particulate matter. J Thromb Haemost 9:1359–1367CrossRefGoogle Scholar
  25. Kreyling WG, Möller W, Semmler-Behnke M, Oberdörster G (2007) Particle dosimetry: deposition and clearance from the respiratory tract and translocation towards extrapulmonary sites. Particle toxicology:2007Google Scholar
  26. Kreyling WG, Semmler M, Erbe F, Mayer P, Takenaka S, Schulz H, Oberdörster G, Ziesenis A (2002) Translocation of ultrafine insoluble iridium particles from lung epithelium to extrapulmonary organs is size dependent but very low. J Toxicol Environ Health A 65:1513–1530. CrossRefGoogle Scholar
  27. Lawal A, Davids L, Marnewick J (2016) Diesel exhaust particles and endothelial cells dysfunction: an update. Toxicol in Vitro 32:92–104CrossRefGoogle Scholar
  28. Li N, Sioutas C, Cho A, Schmitz D, Misra C, Sempf J, Wang M, Oberley T, Froines J, Nel A (2003) Ultrafine particulate pollutants induce oxidative stress and mitochondrial damage. Environ Health Perspect 111:455–460CrossRefGoogle Scholar
  29. Libby P, Ridker PM, Maseri A (2002) Inflammation and atherosclerosis. Circulation 105:1135–1143CrossRefGoogle Scholar
  30. Meier R, Cascio WE, Ghio AJ, Wild P, Danuser B, Riediker M (2014) Associations of short-term particle and noise exposures with markers of cardiovascular and respiratory health among highway maintenance workers. Environ Health Perspect 122:726–732CrossRefGoogle Scholar
  31. Nemmar A, Hoet PH, Vermylen J, Nemery B, Hoylaerts MF (2004) Pharmacological stabilization of mast cells abrogates late thrombotic events induced by diesel exhaust particles in hamsters. Circulation 110:1670–1677CrossRefGoogle Scholar
  32. Nemmar A, Hoet PHM, Vanquickenborne B, Dinsdale D, Thomeer M, Hoylaerts MF, Vanbilloen H, Mortelmans L, Nemery B (2002) Passage of inhaled particles into the blood circulation in humans. Circulation 105:411–414CrossRefGoogle Scholar
  33. Nemmar A, Nemery B, Hoet PH, Van Rooijen N, Hoylaerts MF (2005) Silica particles enhance peripheral thrombosis: key role of lung macrophage–neutrophil cross-talk. Am J Respir Crit Care Med 171:872–879CrossRefGoogle Scholar
  34. Nemmar A, Nemery B, Hoet PH, Vermylen J, Hoylaerts MF (2003) Pulmonary inflammation and thrombogenicity caused by diesel particles in hamsters: role of histamine. Am J Respir Crit Care Med 168:1366–1372CrossRefGoogle Scholar
  35. Nemmar A, Vanbilloen H, Hoylaerts M, Hoet P, Verbruggen A, Nemery B (2001) Passage of intratracheally instilled ultrafine particles from the lung into the systemic circulation in hamster. Am J Respir Crit Care Med 164:1665–1668CrossRefGoogle Scholar
  36. Niemann B, Rohrbach S, Miller MR, Newby DE, Fuster V, Kovacic JC (2017) Oxidative stress and cardiovascular risk: obesity, diabetes, smoking, and pollution: part 3 of a 3-part series. J Am Coll Cardiol 70:230–251CrossRefGoogle Scholar
  37. Oberdörster G, Oberdörster E, Oberdörster J (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 113:823–839CrossRefGoogle Scholar
  38. Oberdörster G, Sharp Z, Atudorei V, Elder A, Gelein R, Lunts A, Kreyling W, Cox C (2002) Extrapulmonary translocation of ultrafine carbon particles following whole-body inhalation exposure of rats. J Toxicol Environ Health, Part A 65:1531–1543CrossRefGoogle Scholar
  39. Oberdörster G, Utell MJ (2002) Ultrafine particles in the urban air: to the respiratory tract--and beyond? Environ Health Perspect 110:A440–a441CrossRefGoogle Scholar
  40. Pan X, Gong YY, Xu Y, Ariens RA, Routledge MN (2018) Urban particulate matter induces changes in gene expression in vascular endothelial cells that are associated with altered clot structure in vitro. Thromb Haemost 118:266–278CrossRefGoogle Scholar
  41. Requia WJ, Adams MD, Arain A, Papatheodorou S, Koutrakis P, Mahmoud M (2018) Global association of air pollution and cardiorespiratory diseases: a systematic review, meta-analysis, and investigation of modifier variables. Am J Public Health 108:S123–S130CrossRefGoogle Scholar
  42. Rodriguez-Yanez Y, Bahena-Uribe D, Chavez-Munguia B, López-Marure R, Gonzalez-Monroy S, Cisneros B, Albores A (2015) Commercial single-walled carbon nanotubes effects in fibrinolysis of human umbilical vein endothelial cells. Toxicol in Vitro 29:1201–1214CrossRefGoogle Scholar
  43. Rondaij MG, Bierings R, Kragt A, van Mourik JA, Voorberg J (2006) Dynamics and plasticity of Weibel-Palade bodies in endothelial cells. Arterioscler Thromb Vasc Biol 26:1002–1007CrossRefGoogle Scholar
  44. Rückerl R et al (2007) Ultrafine particles and platelet activation in patients with coronary heart disease – results from a prospective panel study. Part Fibre Toxicol 4(1):1CrossRefGoogle Scholar
  45. Sahu D, Kannan G, Vijayaraghavan R (2014) Carbon black particle exhibits size dependent toxicity in human monocytes. Int J Inflamm 2014:1–10CrossRefGoogle Scholar
  46. Schmid O, Möller W, Semmler-Behnke M, A. Ferron G, Karg E, Lipka J, Schulz H, Kreyling WG, Stoeger T (2009) Dosimetry and toxicology of inhaled ultrafine particles. Biomarkers 14:67–73CrossRefGoogle Scholar
  47. Schwartz J, Dockery DW, Neas LM (1996) Is daily mortality associated specifically with fine particles? J Air Waste Manage Assoc 46:927–939CrossRefGoogle Scholar
  48. Seagrave J (2008) Mechanisms and implications of air pollution particle associations with chemokines. Toxicol Appl Pharmacol 232:469–477. CrossRefGoogle Scholar
  49. Shah ASV, Langrish JP, Nair H, McAllister DA, Hunter AL, Donaldson K, Newby DE, Mills NL (2013) Global association of air pollution and heart failure: a systematic review and meta-analysis. Lancet 382:1039–1048. CrossRefGoogle Scholar
  50. Shahani T, Covens K, Lavend'Homme R, Jazouli N, Sokal E, Peerlinck K, Jacquemin M (2014) Human liver sinusoidal endothelial cells but not hepatocytes contain factor VIII. J Thromb Haemost 12:36–42CrossRefGoogle Scholar
  51. Shahani T, Lavend'homme R, Luttun A, Saint-Remy JM, Peerlinck K, Jacquemin M (2010) Activation of human endothelial cells from specific vascular beds induces the release of a FVIII storage pool. Blood 115:4902–4909. CrossRefGoogle Scholar
  52. Shrey K, Suchit A, Deepika D, Shruti K, Vibha R (2011) Air pollutants: the key stages in the pathway towards the development of cardiovascular disorders. Environ Toxicol Pharmacol 31:1–9. CrossRefGoogle Scholar
  53. Silverman RA, Ito K, Freese J, Kaufman BJ, De Claro D, Braun J, Prezant DJ (2010) Association of ambient fine particles with out-of-hospital cardiac arrests in New York City. Am J Epidemiol 172:917–923CrossRefGoogle Scholar
  54. Steinvil A, Kordova-Biezuner L, Shapira I, Berliner S, Rogowski O (2008) Short-term exposure to air pollution and inflammation-sensitive biomarkers. Environ Res 106:51–61. CrossRefGoogle Scholar
  55. Burnett RT et al (2000) Association between particulate-and gas-phase components of urban air pollution and daily mortality in eight Canadian cities. Inhal Toxicol 12:15–39CrossRefGoogle Scholar
  56. Thurston GD, Burnett RT, Turner MC, Shi Y, Krewski D, Lall R, Ito K, Jerrett M, Gapstur SM, Diver WR, Pope CA III (2016) Ischemic heart disease mortality and long-term exposure to source-related components of US fine particle air pollution. Environ Health Perspect 124:785–794CrossRefGoogle Scholar
  57. Valentijn K, Valentijn J, Jansen K, Koster A (2008) A new look at Weibel–Palade body structure in endothelial cells using electron tomography. J Struct Biol 161:447–458CrossRefGoogle Scholar
  58. Valentijn KM, Sadler JE, Valentijn JA, Voorberg J, Eikenboom J (2011) Functional architecture of Weibel-Palade bodies. Blood 117:5033–5043CrossRefGoogle Scholar
  59. van Eeden SF et al (2001) Cytokines involved in the systemic inflammatory response induced by exposure to particulate matter air pollutants (PM(10)). Am J Respir Crit Care Med 164:826–830. CrossRefGoogle Scholar
  60. Vesterdal LK, Mikkelsen L, Folkmann JK, Sheykhzade M, Cao Y, Roursgaard M, Loft S, Møller P (2012) Carbon black nanoparticles and vascular dysfunction in cultured endothelial cells and artery segments. Toxicol Lett 214:19–26CrossRefGoogle Scholar
  61. Walker VG, Li Z, Hulderman T, Schwegler-Berry D, Kashon ML, Simeonova PP (2009) Potential in vitro effects of carbon nanotubes on human aortic endothelial cells. Toxicol Appl Pharmacol 236:319–328CrossRefGoogle Scholar
  62. Wang J, Tang B, Liu X, Wu X, Wang H, Xu D, Guo Y (2015) Increased monomeric CRP levels in acute myocardial infarction: a possible new and specific biomarker for diagnosis and severity assessment of disease. Atherosclerosis 239:343–349. CrossRefGoogle Scholar
  63. Yamagami H, Yamagami S, Inoki T, Amano S, Miyata K (2003) The effects of proinflammatory cytokines on cytokine-chemokine gene expression profiles in the human corneal endothelium. Invest Ophthalmol Vis Sci 44:514–520CrossRefGoogle Scholar
  64. Yamawaki H, Iwai N (2006) Mechanisms underlying Nano-sized air-pollution-mediated progression of atherosclerosis. Circ J 70:129–140CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Genetics and Molecular Medicine, School of MedicineZanjan University of Medical Sciences (ZUMS)ZanjanIran
  2. 2.Department of Cardiovascular SciencesCenter for Molecular and Vascular Biology, KU LeuvenLeuvenBelgium
  3. 3.Department of Anatomy, School of MedicineZanjan University of Medical Sciences (ZUMS)ZanjanIran

Personalised recommendations