, Volume 31, Issue 1, pp 65-79

Nitric Oxide Delivery System for Cell Culture Studies

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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 NO 2 (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 NO 2 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.