Cholesterol Modifies the Properties of Surface Films of Dipalmitoylphosphatidylcholine plus Pulmonary Surfactant-Associated Protein B or C Spread or Adsorbed at the Air−Water Interface

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Abstract
Cholesterol is a substantial component of pulmonary surfactant (approximately 8 wt % or approximately 14 mol % of surfactant lipids). This study investigated the effect of cholesterol on the way in which hydrophobic SP-B and SP-C modulated the adsorption of lipid into the air-water interface and their respreading from collapsed phase produced on overcompression of the surface film. The properties of binary spread monolayers of SP-B or SP-C plus cholesterol (CH) were consistent with miscibility between the hydrophobic proteins and the sterol. Results from surface pressure versus area measurements at 23 degrees C on spread monolayers of dipalmitoylphosphatidylcholine (DPPC) plus SP-B in the presence of 8 wt % cholesterol implied that CH did not significantly affect the properties of the films of SP-B/(DPPC/CH) compared to those of binary SP-B/DPPC monolayers. In contrast, CH appeared to enhance the mixing of SP-C with DPPC/CH in ternary SP-C/(DPPC/CH) films compared to the miscibility of SP-C with DPPC in the SP-C/DPPC films. It is estimated that about 10 wt % SP-C might remain in the SP-C/(DPPC/CH) monolayers compressed to high surface pressures of about 72 mN/m, whereas SP-C at concentrations of > or = 5 wt % was squeezed out at pi approximately 50 mN/m from SP-C/DPPC films without cholesterol. Cholesterol reduced the stability of the films of SP-B/(DPPC/CH) and SP-C/(DPPC/CH) when they had been compressed to pi approximately 72 mN/m, in contrast to films of SP-B/DPPC and SP-C/DPPC which exhibited a relatively slow relaxation from the collapse pressure of 72 mN/m. Dynamic cyclic compression beyond collapse of SP-B/(DPPC/CH) and SP-C/(DPPC/CH) monolayers showed that cholesterol diminished their postcollapse respreading compared to the respreading of the protein/DPPC films without cholesterol. Cholesterol, at 8 wt %, inhibited the rate of adsorption to the air-water interface at 35 degrees C of aqueous dispersions of DPPC containing 2.5 or 5 wt % SP-B or SP-C. The results suggest that cholesterol has an apparent negative influence on the surfactant surface properties, which are generally considered to be important in surfactant function, although increasing protein concentrations can counteract some of the negative influences.