Abstract
BackgroundCystic fibrosis (CF) is a life-threatening multiorgan genetic disease, particularly affecting the lungs, where recurrent infections are the main cause of reduced life expectancy. In CF, mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein impair transepithelial electrolyte and water transport, resulting in airway dehydration, and a thickening of the mucus associated with abnormal viscoelastic properties. Our aim was to develop a rheological method to assess the effects of hypertonic saline (NaCl) and NaHCO3 on CF sputum viscoelasticity in vitro, and to identify the critical steps in sample preparation and in the rheological measurements.MethodsSputum samples were mixed with hypertonic salt solutions in vitro in a ratio of either 10:4 or 10:1. Distilled water was applied as a reference treatment. The rheological properties of sputum from CF patients, and the effects of these in vitro treatments, were studied with a rheometer at constant frequency and strain, followed by frequency sweep tests, where storage modulus (G′), loss modulus (G″) and loss factor were determined.ResultsWe identified three distinct categories of sputum: (i) highly elastic (G′ > 100,000 Pa), (ii) elastic (100,000 Pa > G′ > 1000 Pa), and (iii) viscoelastic (G′ < 1000). At the higher additive ratio (10:4), all of the added solutions were found to significantly reduce the gel strength of the sputum, but the most pronounced changes were observed with NaHCO3 (p < 0.001). Samples with high elasticity exhibited the greatest changes while, for less elastic samples, a weakening of the gel structure was observed when they were treated with water or NaHCO3, but not with NaCl. For the viscoelastic samples, the additives did not cause significant changes in the parameters. When the lower additive ratio (10:1) was used, the mean values of the rheological parameters usually decreased, but the changes were not statistically significant.ConclusionBased on the rheological properties of the initial sputum samples, we can predict with some confidence the treatment efficacy of each of the alternative additives. The marked differences between the three categories suggest that it is advisable to evaluate each sample individually using a rheological approach such as that described here.
Highlights
Cystic fibrosis (CF) is one of the most common lifethreatening genetic diseases
In order to analyse the possible breakdown of mucus structure in CF, and to follow the recovery of the sputum samples after their treatment and insertion into the rheometer, a constant oscillation test was applied at low frequency and strain value
In order to distinguish between these two possibilities, experiments were performed with sputum samples pre-incubated at 37 °C for 30 min and inserted into the pre-warmed instrument at the same temperature
Summary
Cystic fibrosis (CF) is one of the most common lifethreatening genetic diseases. It affects many organs, including the lungs, where viscous mucus clogs the airways and recurrent infections shorten the patients’Budai‐Szűcs et al BMC Pulm Med (2021) 21:225 lifespan [1]. Cystic fibrosis (CF) is one of the most common lifethreatening genetic diseases It affects many organs, including the lungs, where viscous mucus clogs the airways and recurrent infections shorten the patients’. The rheological properties of the mucus strongly influence the effectiveness of mucus clearance and cough [2,3,4] Changes in these properties significantly reduce mucus clearance in CF and contribute to the colonization of the airways by bacteria and the development of recurrent infections [5, 6]. In CF, mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein impair transepithelial electrolyte and water transport, resulting in airway dehydration, and a thickening of the mucus associated with abnormal viscoelastic properties. Our aim was to develop a rheological method to assess the effects of hypertonic saline (NaCl) and N aHCO3 on CF sputum viscoelasticity in vitro, and to identify the critical steps in sample preparation and in the rheological measurements
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