Comparison of methods for the analysis of airway macrophage particulate load from induced sputum, a potential biomarker of air pollution exposure

BMC Pulm Med. 2015 Nov 5:15:137. doi: 10.1186/s12890-015-0135-7.

Abstract

Background: Air pollution is associated with a high burden or morbidity and mortality, but exposure cannot be quantified rapidly or cheaply. The particulate burden of macrophages from induced sputum may provide a biomarker. We compare the feasibility of two methods for digital quantification of airway macrophage particulate load.

Methods: Induced sputum samples were processed and analysed using ImageJ and Image SXM software packages. We compare each package by resources and time required.

Results: 13 adequate samples were obtained from 21 patients. Median particulate load was 0.38 μm(2) (ImageJ) and 4.0 % of the total cellular area of macrophages (Image SXM), with no correlation between results obtained using the two methods (correlation coefficient = -0.42, p = 0.256). Image SXM took longer than ImageJ (median 26 vs 54 mins per participant, p = 0.008) and was less accurate based on visual assessment of the output images. ImageJ's method is subjective and requires well-trained staff.

Conclusion: Induced sputum has limited application as a screening tool due to the resources required. Limitations of both methods compared here were found: the heterogeneity of induced sputum appearances makes automated image analysis challenging. Further work should refine methodologies and assess inter- and intra-observer reliability, if these methods are to be developed for investigating the relationship of particulate and inflammatory response in the macrophage.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Air Pollution*
  • Asthma / physiopathology*
  • Biomarkers
  • Bronchiectasis / physiopathology*
  • Environmental Exposure
  • Feasibility Studies
  • Female
  • Forced Expiratory Volume
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Inhalation Exposure*
  • Macrophages, Alveolar / pathology*
  • Male
  • Middle Aged
  • Particulate Matter / analysis*
  • Reproducibility of Results
  • Software
  • Sputum / cytology*
  • Vital Capacity

Substances

  • Biomarkers
  • Particulate Matter