The Effect of Cholesterol on the Biophysical Inhibition of Pulmonary Surfactant by Albumin

Abstract

Pulmonary surfactant is a mixture of 85% phospholipids, 5–8% cholesterol and 5–8% surfactant proteins which forms a surface tension reducing lipid film at the air‐liquid interface of the alveoli. By reducing the surface tension to near 0mN/m values during compression (i.e. exhalation) surfactant maintains alveolar stability at low lung volumes and maintains proper lung compliance. The importance of surfactant is evident in Acute Respiratory Distress Syndrome (ARDS) in which the dysfunction of surfactant contributes to hypoxemia, regional areas of collapse and reduced lung compliance. Specifically, serum proteins, such as Albumin, leaking into the lung during ARDS have been shown to interfere with surfactant's ability to reach low surface tension. In addition, recent studies suggest that elevated cholesterol within surfactant also contribute to surfactant dysfunction in the setting of lung injury. Although the effects of cholesterol and serum proteins with surfactant have been studied independently, the interaction between the two has not previously been investigated. It was hypothesized that elevated levels of cholesterol within surfactant will make surfactant more susceptible to serum protein inhibition.MethodsBovine Lipid Extract Surfactant (BLES), a commercially available exogenous surfactant, was utilized and modified to generate samples with concentrations of 0, 2.5, 5 and 10% (w/w) cholesterol. Varying amounts of Bovine Serum Albumin (BSA) were subsequently added to each sample to attain concentrations of 0, 20, 30, 40 and 50 mg/ml. A constrained sessile drop surfactometer was used to determine the minimal achievable surface tensions during dynamic compression/expansion cycles.ResultsSamples with various levels of cholesterol, in the absence of BSA, reached minimum surface tension values of near 0mN/m during compression. At moderate levels of BSA (20 & 30mg/ml), minimum surface tensions were lower in surfactant samples containing cholesterol as compared to samples containing no cholesterol. Finally, at high BSA values, all surfactant samples had surface tension values that were higher than those at the lower BSA concentration but were not significantly different among the sample with different cholesterol concentrations.DiscussionContrary to the hypothesis, values of up to 10% cholesterol within surfactant do not appear to increase the susceptibility of surfactant to serum protein inhibition. In fact, the results suggest that cholesterol mitigates the effect of moderate levels of BSA on surfactant function. It is concluded that cholesterol within surfactant contributes to its resistance against inhibition by albumin.Support or Funding InformationCanadian Institutes for Health Research.