Annals of Biomedical Engineering

, Volume 31, Issue 1, pp 65–79 | Cite as

Nitric Oxide Delivery System for Cell Culture Studies

  • Chen Wang
  • William M. Deen
Article

Abstract

To investigate the toxicity and mutagenicity of NO, methods are needed to deliver it to cell cultures at known, constant rates. To permit continuous exposures over lengthy periods, we fabricated a simple apparatus utilizing gas-permeable polydimethylsiloxane (Silastic) tubing to supply both NO and O2 to a stirred, cylindrical vessel. Mass transfer in this system was characterized by measuring the delivery rates of NO or O2 alone, and of NO to air-saturated solutions. The concentrations of NO, O2 and NO2 (the end product of NO oxidation) were monitored continuously. The total flux of nitrogen species into the liquid (as determined from the sum of NO and NO2 accumulation) was 50%–90% greater in the presence of O2 depending on the NO partial pressure in the gas. Also, the simultaneously measured mass transfer coefficients for NO and O2 differed greatly from the corresponding unreactive values. An analysis of the data using diffusion-reaction models showed that NO oxidation in the aqueous boundary layer contributed very little to the nitrogen flux increase or to variations in the mass transfer coefficients. However, the unusually strong dependence of the delivery rates on chemical reactions could be explained by postulating that partial oxidation of NO to NO2 occurred within the membrane. The rate constant we estimated for polydimethylsiloxane, >4.4 × 105 M−2 s−1 at 23°C, is only about one-fifth of values reported previously for water and nonpolar solvents, but the high solubilities of NO and O2 in the polymer are sufficient to make NO2 formation significant. Although considerable NO2 is calculated to enter the liquid, its reaction with aqueous NO is rapid enough to keep this undesired compound at trace levels, except within a few microns of the tubing. Thus, cells will have little exposure to NO2. © 2003 Biomedical Engineering Society.

Nitric oxide, Toxicity of Polydimethylsiloxane, Gas diffusion in Silastic, Gas diffusion in Gas-liquid mass transfer 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

REFERENCES

  1. 1.
    Buga, G. M., and L. J. Ignarro. Nitric oxide and cancer. In: Nitric Oxide Physiology and Pathology, edited by L. J. Ignarro. San Diego: Academic Press, 2000, pp. 895–920.­Google Scholar
  2. 2.
    Deen, W. M. Analysis of Transport Phenomena. New York: Oxford University Press, 1998, pp. 422–429.­Google Scholar
  3. 3.
    Estevez, A. G., N. Spear, H. Pelluffo, A. Kamaid, L. Barbeito, and J. S. Beckman. Examining apoptosis in cultured cells after exposure to nitric oxide and peroxynitrite. Methods Enzymol.301:393­402, 1999.­Google Scholar
  4. 4.
    Gasco, A., R. Fruttero, and G. Sorba. NO Donors: An emerging class of compounds in medicinal chemistry. Farmacognosia51:617­635, 1996.­Google Scholar
  5. 5.
    Graetzel, M., S. Taniguchi, and A. Henglein. Pulse radiolytic investigation of NO oxidation and equilibrium N2O3=NO+NO2 in aqueous solution. Ber. Bunsenges. Phys. Chem.74:488­492, 1970.­Google Scholar
  6. 6.
    Green, L. C., D. A. Wagner, J. Glogowski, P. L. Skipper, and J. S. Wishnok. Analysis of nitrate, nitrite, and 15NO3 in biological fluids. Anal. Biochem.126:131­138, 1982.­Google Scholar
  7. 7.
    Kavdia, M., S. Nagarajan, and R. S. Lewis. Novel devices for the predictable delivery of nitric oxide to aqueous solutions. Chem. Res. Toxicol.11:1346­1351, 1998.­Google Scholar
  8. 8.
    Kavdia, M., J. L. Stanfield, and R. S. Lewis. Nitric oxide, superoxide, and peroxynitrite effects on the insulin secretion and viability of βTC3 cells. Ann. Biomed. Eng.28:102­109, 2000.­Google Scholar
  9. 9.
    Lange's Handbook of Chemistry, Rev. 10th ed., edited by N. A. Lange. New York: McGraw­Hill, 1967.­Google Scholar
  10. 10.
    Lewis, R. S., and W. M. Deen. Kinetics of the reaction of nitric oxide with oxygen in aqueous solutions. Chem. Res. Toxicol.7:568­574, 1994.­Google Scholar
  11. 11.
    Lewis, R. S., W. M. Deen, S. R. Tannenbaum, and J. S. Wishnok. Membrane mass spectrometer inlet for quantitation of nitric oxide. Biol. Mass Spectrom.22:45­52, 1993.­Google Scholar
  12. 12.
    Lewis, R. S., S. Tamir, and W. M. Deen. Kinetic analysis of the fate of nitric oxide synthesized by macrophages. J. Biol. Chem.270:29350­29355, 1995.­Google Scholar
  13. 13.
    Licht, W. R., S. R. Tannenbaum, and W. M. Deen. Use of ascorbic acid to inhibit nitrosation: Kinetic and mass transfer considerations for an system. Carcinogenesis9:365­372, 1988.­Google Scholar
  14. 14.
    Liu, X., M. J. S. Miller, M. S. Joshi, D. D. Thomas, and J. R. Lancaster. Accelerated reaction of nitric oxide with O2 within the hydrophobic interior of biological membranes. Proc. Natl. Acad. Sci. U.S.A.95:2175­2179, 1998.­Google Scholar
  15. 15.
    Mertes, S., and A. Wahner. Uptake of nitrogen dioxides and nitrous acid on aqueous surfaces. J. Phys. Chem.99:14000­14006, 1995.­Google Scholar
  16. 16.
    Moncada, S., R. M. J. Palmer, and E. A. Higgs. Nitric oxide: Physiology, pathophysiology and pharmacology. Pharmacol. Rev.43:109­142, 1991.­Google Scholar
  17. 17.
    Nottingham, W., and J. R. Sutter. Kinetics of the oxidation of nitric oxide by chlorine and oxygen in nonaqueous media. Int. J. Chem. Kinet.18:1289­1302, 1986.­Google Scholar
  18. 18.
    Patel, R. P., and V. M. Darley­Usmar. Using peroxynitrite as oxidant with low­density lipoprotein. Methods Enzymol.269:375­384, 1996.­Google Scholar
  19. 19.
    Perry's Chemical Engineer's Handbook, 6th ed., edited by Perry, R. H. and D. W. Green. New York: McGraw­Hill, 1984, p. 3­252.­Google Scholar
  20. 20.
    Pfeiffer, S., A. Lass, K. Schmidt, and B. Mayer. Protein tyrosine nitration in mouse peritoneal macrophages activated and: Evidence against an essential role of peroxynitrite. FASEB J.15:2355­2364, 2001.­Google Scholar
  21. 21.
    Press, W. H., S. A. Teukolsky, W. T. Vetterling, and B. P. Flannery. Numerical Recipes in FORTRAN: The Art of Scientific Computing, 2nd ed. Cambridge: Cambridge University Press, 1992, pp. 704–707, 749–752.­Google Scholar
  22. 22.
    Ramamurthi, A., and R. S. Lewis. Measurement and modeling of nitric oxide release rates for nitric oxide donors. Chem. Res. Toxicol.10:408­413, 1997.­Google Scholar
  23. 23.
    Ramamurthi, A., and R. S. Lewis. Design of a novel apparatus to study nitric oxide (NO) inhibition of platelet adhesion. Ann. Biomed. Eng.26:1036­1043, 1998.­Google Scholar
  24. 24.
    Ramamurthi, A., and R. S. Lewis. Influence of agonist, shear rate, and perfusion time on nitric oxide inhibition of platelet deposition. Ann. Biomed. Eng.28:174­181, 2000.­Google Scholar
  25. 25.
    Robb, W. L.Thin silicone membranes—Their permeation properties and some applications. Ann. N.Y. Acad. Sci.46:119­137, 1968.­Google Scholar
  26. 26.
    Schmidt, K., and B. Mayer. Determination of NO with a Clark­type electrode. In: Nitric Oxide Protocols, edited by M. A. Titheradge. Totowa, NJ: Humana, 1998, pp. 101–109.­Google Scholar
  27. 27.
    Schumpe, A.The estimation of gas solubility in salt solutions. Chem. Eng. Sci.48:153­158, 1993.­Google Scholar
  28. 28.
    Schwartz, S. E. Trace Atmosphere Constituents: Properties, Transformations, and Fates. New York: Wiley, 1983.­Google Scholar
  29. 29.
    Tamir, S., R. S. Lewis, T. D. R. Walker, W. M. Deen, J. S. Wishnok, and S. T. Tannenbaum. The influence of delivery rate on the chemistry and biological effects of nitric oxide. Chem. Res. Toxicol.6:895­899, 1993.­Google Scholar
  30. 30.
    Thomas, D. D., X. Liu, S. P. Kantrow, and J. R. Lancaster. The biological life of nitric oxide: Implications for the perivascular dynamics of NO and O2.Proc. Natl. Acad. Sci. U.S.A.98:355­360, 2001.­Google Scholar
  31. 31.
    Treacy, J. C., and F. Daniels. Kinetic studies of the oxidation of nitric oxide with oxygen in the pressure range 1 to 20 Mm. J. Am. Chem. Soc.77:2033­2035, 1955.­Google Scholar
  32. 32.
    Weisenberger, S., and A. Schumpe. Estimation of gas solubility in salt solutions at temperatures from 273 K to 363 K. AIChE J.42:298­300, 1996.­Google Scholar
  33. 33.
    Wilke, C. P., and P. Chang. Correlation of diffusion coefficient in dilute solutions. AIChE J.2:264­270, 1955.­Google Scholar
  34. 34.
    Wise, D. L., and G. Houghton. Diffusion coefficients of neon, krypton, xenon, carbon monooxide, and nitric oxide in water at 10–60°C. Chem. Eng. Sci.23:1211­1216, 1968.­ ­Google Scholar

Copyright information

© Biomedical Engineering Society 2003

Authors and Affiliations

  • Chen Wang
    • 1
  • William M. Deen
    • 2
    • 3
  1. 1.Department of Chemical EngineeringMassachusetts Institute of TechnologyCambridge
  2. 2.Department of Chemical EngineeringUSA
  3. 3.Biological Engineering DivisionMassachusetts Institute of TechnologyCambridge

Personalised recommendations