Platelet and leukocyte adhesion to albumin binding self-assembled monolayers

  • Inês C. Gonçalves
  • M. Cristina L. Martins
  • Judite N. Barbosa
  • Pedro Oliveira
  • Mário A. Barbosa
  • Buddy D. Ratner


This study reports the use of tetraethylene glycol-terminated self-assembled monolayers (EG4 SAMs) as a background non-fouling surface to study the effect of an 18 carbon ligand (C18) on albumin selective and reversible adsorption and subsequent platelet and leukocyte adhesion. Surface characterization techniques revealed an efficient immobilization of different levels of C18 ligand on EG4 SAMs and an increase of surface thickness and hydrophobicity with the increase of C18 ligands. Albumin adsorption increased as the percentage of C18 ligands on the surface increased, but only 2.5%C18 SAMs adsorbed albumin in a selective and reversible way. Adherent platelets also increased with the amount of immobilized C18. Pre-immersion of samples in albumin before contact with platelets demonstrated an 80% decrease in platelet adhesion. Pre-immersion in plasma was only relevant for 2.5%C18 SAMs since this was the only surface to have less platelet adhesion compared to buffer pre-immersion. EG4 SAMs adhered negligible amounts of leukocytes, but surfaces with C18 ligands have some adherent leukocytes. Except for 10%C18 SAMs, which increased leukocyte adhesion after albumin pre-adhesion, protein pre-immersion did not influence leukocyte adhesion. It has been shown that a surface with a specific surface concentration of albumin-binding ligands (2.5%C18 SAMs) can recruit albumin selectively and reversibly and minimize the adhesion of platelets, despite still adhering some leukocytes.


  1. 1.
    Gorbet MB, Sefton MV. Biomaterial-associated thrombosis: roles of coagulation factors, complement, platelets and leukocytes. Biomaterials. 2004;25(26):5681–703.CrossRefGoogle Scholar
  2. 2.
    Lindblad M, et al. Cell and soft tissue interactions with methyl- and hydroxyl-terminated alkane thiols on gold surfaces. Biomaterials. 1997;18(15):1059–68.CrossRefGoogle Scholar
  3. 3.
    Anderson JM. Biological responses to materials. Annu Rev Mater Res. 2001;31:81–110.CrossRefGoogle Scholar
  4. 4.
    Mao C, et al. Various approaches to modify biomaterial surfaces for improving hemocompatibility. Adv Colloid Interface Sci. 2004;110(1–2):5–17.CrossRefGoogle Scholar
  5. 5.
    Barbosa JN, et al. Adhesion of human leukocytes on mixtures of hydroxyl- and methyl-terminated self-assembled monolayers: effect of blood protein adsorption. J Biomed Mater Res A. 2010;93A(1):12–9.Google Scholar
  6. 6.
    Collier TO, Anderson JM. Protein and surface effects on monocyte and macrophage adhesion, maturation, and survival. J Biomed Mater Res. 2002;60(3):487–96.CrossRefGoogle Scholar
  7. 7.
    Jenney CR, Anderson JM. Adsorbed serum proteins responsible for surface dependent human macrophage behavior. J Biomed Mater Res. 1999;49(4):435–47.CrossRefGoogle Scholar
  8. 8.
    Nimeri G, et al. The influence of plasma proteins and platelets on oxygen radical production and F-actin distribution in neutrophils adhering to polymer surfaces. Biomaterials. 2002;23(8):1785–95.CrossRefGoogle Scholar
  9. 9.
    Tidwell CD, et al. Endothelial cell growth and protein adsorption on terminally functionalized, self-assembled monolayers of alkanethiolates on gold. Langmuir. 1997;13(13):3404–13.CrossRefGoogle Scholar
  10. 10.
    Lee HB, Kim SS, Khang G. Polymeric biomaterials. In: Bronzino JD, editor. The biomedical engineering handbook. Boca Raton: CRC Press; 1995. p. 581–97.Google Scholar
  11. 11.
    Keogh JR, Eaton JW. Albumin-binding surfaces for biomaterials. J Lab Clin Med. 1994;124(4):537–45.Google Scholar
  12. 12.
    Kottkemarchant K, et al. Effect of albumin coating on the invitro blood compatibility of dacron arterial prostheses. Biomaterials. 1989;10(3):147–55.CrossRefGoogle Scholar
  13. 13.
    Ji J, Feng LX, Barbosa MA. Stearyl poly(ethylene oxide) grafted surfaces for preferential adsorption of albumin. Biomaterials. 2001;22(22):3015–23.CrossRefGoogle Scholar
  14. 14.
    Munro MS, et al. Alkyl substituted polymers with enhanced albumin affinity. Trans Am Soc Artif Intern Organs. 1981;27:499–503.Google Scholar
  15. 15.
    Martins MCL, et al. Albumin adsorption on Cibacron Blue F3G-A immobilized onto oligo(ethylene glycol)- terminated self-assembled monolayers. J Mater Sci. 2003;14:945–54.CrossRefGoogle Scholar
  16. 16.
    Martins MCL, et al. Albumin and fibrinogen adsorption on Cibacron blue F3G-A immobilised onto PU-PHEMA (polyurethane-poly(hydroxyethylmethacrylate)) surfaces. J Biomater Sci. 2003;14(5):439–55.CrossRefGoogle Scholar
  17. 17.
    Ji J, et al. Preparation of albumin preferential surfaces on poly(vinyl chloride) membranes via surface self-segregation. J Biomed Mater Res. 2002;61(2):252–9.CrossRefGoogle Scholar
  18. 18.
    Guha Thakurta S, Subramanian A. Evaluation of in situ albumin binding surfaces: a study of protein adsorption and platelet adhesion. J Mater Sci Mater Med. 2011;22(1):137–49.CrossRefGoogle Scholar
  19. 19.
    McFarland CD, et al. Albumin-binding surfaces: in vitro activity. J Biomater Sci. 1998;9(11):1227–39.CrossRefGoogle Scholar
  20. 20.
    Goncalves IC, et al. Protein adsorption on 18-alkyl chains immobilized on hydroxyl-terminated self-assembled monolayers. Biomaterials. 2005;26(18):3891–9.CrossRefGoogle Scholar
  21. 21.
    Gonçalves IC, et al. Selective protein adsorption modulates platelet adhesion and activation to oligo(ethylene glycol)-terminated self-assembled monolayers with C18 ligands. J Biomed Mater Res A. 2009;89A(3):642–53.CrossRefGoogle Scholar
  22. 22.
    Owens DK, Wendt RC. Estimation of surface free energy of polymers. J Appl Polym Sci. 1969;13(8):1741–7.CrossRefGoogle Scholar
  23. 23.
    Sousa SR, et al. Human serum albumin adsorption on TiO2 from single protein solutions and from plasma. Langmuir. 2004;20:9745–54.CrossRefGoogle Scholar
  24. 24.
    Sastry M. A note on the use of ellipsometry for studying the kinetics of formation of self-assembled monolayers. Bull Mater Sci. 2000;23(3):159–63.CrossRefGoogle Scholar
  25. 25.
    Rodrigues SN, et al. Fibrinogen adsorption, platelet adhesion and activation on mixed hydroxyl-/methyl-terminated self-assembled monolayers. Biomaterials. 2006;27(31):5357–67.CrossRefGoogle Scholar
  26. 26.
    Tang LP, et al. Molecular determinants of acute inflammatory responses to biomaterials. J Clin Invest. 1996;97(5):1329–34.CrossRefGoogle Scholar
  27. 27.
    Martins MCL, Ratner BD, Barbosa MA. Protein adsorption on mixtures of hydroxyl- and methylterminated alkanethiols self-assembled monolavers. J Biomed Mater Res A. 2003;67A(1):158–71.CrossRefGoogle Scholar
  28. 28.
    Bain CD, et al. Formation of monolayer films by the spontaneous assembly of organic thiols from solution onto gold. J Am Chem Soc. 1989;111(1):321–35.CrossRefGoogle Scholar
  29. 29.
    Palegrosdemange C, et al. Formation of self-assembled monolayers by chemisorption of derivatives of oligo(ethylene glycol) of structure Hs(Ch2)11(Och2ch2)meta-Oh on gold. J Am Chem Soc. 1991;113(1):12–20.CrossRefGoogle Scholar
  30. 30.
    Harder P, et al. Molecular conformation in oligo(ethylene glycol)-terminated self-assembled monolayers on gold and silver surfaces determines their ability to resist protein adsorption. J Phys Chem B. 1998;102(2):426–36.CrossRefGoogle Scholar
  31. 31.
    Zhu B, et al. Chain-length dependence of the protein and cell resistance of oligo(ethylene glycol)-terminated self-assembled monolayers on gold. J Biomed Mater Res. 2001;56(3):406–16.CrossRefGoogle Scholar
  32. 32.
    Tegoulia VA, Cooper SL. Leukocyte adhesion on model surfaces under flow: effects of surface chemistry, protein adsorption, and shear rate. J Biomed Mater Res. 2000;50(3):291–301.CrossRefGoogle Scholar
  33. 33.
    Wang RLC, Kreuzer HJ, Grunze M. Molecular conformation and solvation of oligo(ethylene glycol)-terminated self-assembled monolayers and their resistance to protein adsorption. J Phys Chem B. 1997;101(47):9767–73.CrossRefGoogle Scholar
  34. 34.
    Brodbeck WG, Colton E, Anderson JM. Effects of adsorbed heat labile serum proteins and fibrinogen on adhesion and apoptosis of monocytes/macrophages on biomaterials. J Mater Sci. 2003;14(8):671–5.CrossRefGoogle Scholar
  35. 35.
    Lim F, Cooper SL. Effect of surface hydrophilicity on biomaterial-leukocyte interactions. ASAIO Trans. 1991;37(3):M146–7.Google Scholar
  36. 36.
    Wettero J, Bengtsson T, Tengvall P. Complement activation on immunoglobulin G-coated hydrophobic surfaces enhances the release of oxygen radicals from neutrophils through an actin-dependent mechanism. J Biomed Mater Res. 2000;51(4):742–51.CrossRefGoogle Scholar
  37. 37.
    Sperling C, et al. In vitro hemocompatibility of self-assembled monolayers displaying various functional groups. Biomaterials. 2005;26(33):6547–57.CrossRefGoogle Scholar
  38. 38.
    Barbosa JN, Barbosa MA, Aguas AP. Inflammatory responses and cell adhesion to self-assembled monolayers of alkanethiolates on gold. Biomaterials. 2004;25(13):2557–63.CrossRefGoogle Scholar
  39. 39.
    Lim F, Cooper SL. Effect of sulphonate incorporation on in vitro leucocyte adhesion to polyurethanes. Biomaterials. 1995;16(6):457–66.CrossRefGoogle Scholar
  40. 40.
    van Oss CJ, Gillman CF, Neumann AW. Phagocytic engulfment and cell adhesiveness as cellular surface phenomena. New York: Marcel Dekker Inc; 1975. p. 160.Google Scholar
  41. 41.
    Horbett TA, Klumb LA. Cell culturing: surface aspects and considerations. In: Brash JL, Wojciechowski PW, editors. Interfacial phenomena and bioproducts. New York: Marcel Dekker; 1996. p. 351.Google Scholar
  42. 42.
    Horbett TA. Principles underlying the role of adsorbed plasma proteins in blood interactions with foreign materials. Cardiovasc Pathol. 1993;2:137–48.CrossRefGoogle Scholar
  43. 43.
    Brash JL, Tenhove P. Effect of plasma dilution on adsorption of fibrinogen to solid-surfaces. Thromb Haemost. 1984;51(3):326–30.Google Scholar
  44. 44.
    Brash JL, Tenhove P. Protein adsorption studies on standard polymeric materials. J Biomater Sci. 1993;4(6):591–9.CrossRefGoogle Scholar
  45. 45.
    Werthen M, et al. In vitro study of monocyte viability during the initial adhesion to albumin- and fibrinogen-coated surfaces. Biomaterials. 2001;22(8):827–32.CrossRefGoogle Scholar
  46. 46.
    McNally AK, Anderson JM. Complement C3 participation in monocyte adhesion to different surfaces. Proc Natl Acad Sci USA. 1994;91(21):10119–23.CrossRefGoogle Scholar
  47. 47.
    Altieri DC, Plescia J, Plow EF. The structural motif glycine-190-valine-202 of the fibrinogen-Gamma chain interacts with Cd11b Cd18 integrin (alpha-M-beta-2, mac-1) and promotes leukocyte adhesion. J Biol Chem. 1993;268(3):1847–53.Google Scholar
  48. 48.
    Altieri DC, et al. Structural recognition of a novel fibrinogen gamma-chain sequence(117–133) by intercellular-adhesion molecule-1 mediates leukocyte-endothelium interaction. J Biol Chem. 1995;270(2):696–9.CrossRefGoogle Scholar
  49. 49.
    Smiley ST, King JA, Hancock WW. Fibrinogen stimulates macrophage chemokine secretion through toll-like receptor 4. J Immunol. 2001;167(5):2887–94.Google Scholar
  50. 50.
    Brevig T, et al. The recognition of adsorbed and denatured proteins of different topographies by beta 2 integrins and effects on leukocyte adhesion and activation. Biomaterials. 2005;26(16):3039–53.CrossRefGoogle Scholar
  51. 51.
    Jones DA, Smith CW, McIntire LV. Leucocyte adhesion under flow conditions: principles important in tissue engineering. Biomaterials. 1996;17(3):337–47.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Inês C. Gonçalves
    • 1
    • 2
  • M. Cristina L. Martins
    • 1
  • Judite N. Barbosa
    • 1
    • 3
  • Pedro Oliveira
    • 3
  • Mário A. Barbosa
    • 1
    • 2
    • 3
  • Buddy D. Ratner
    • 4
  1. 1.INEB-Instituto de Engenharia BiomédicaUniversidade do PortoPortoPortugal
  2. 2.Faculdade de EngenhariaUniversidade do PortoPortoPortugal
  3. 3.ICBAS - Instituto de Ciências Biomédicas de Abel SalazarUniversidade do PortoPortoPortugal
  4. 4.Department of Bioengineering and Chemical EngineeringUniversity of WashingtonSeattleUSA

Personalised recommendations