Woolcock Institute of Medical Research, Level 3 Cell Biology Lab, 431 Glebe Point Road, Glebe, New South Wales 2037, Sydney, AustraliabThe University of Sydney, Central Clinical School, Faculty of Medicine, New South Wales 2006, Sydney, Australia.
The University of Sydney, Australian Centre for Microscopy and Microanalysis, Madsen Building F09, New South Wales 2006, Sydney, Australia.
J Biomed Opt. 2014 Mar;19(3):36005. doi: 10.1117/1.JBO.19.3.036005.
Extracellular matrix (ECM) remodeling contributes to the pathogenic changes in chronic obstructive pulmonary disease (COPD) and is both complex and not well understood. Collagen I, a component of the ECM altered in COPD airways, has second harmonic generation (SHG) properties. The SHG signal is coherent, propagating both forward (F) (primarily organized/mature collagen fibrils) and backward (B) (primarily disorganized/immature collagen fibrils) parallel to the incident light. The F/B SHG ratio was used to determine the proportion of organized to disorganized collagen, with lower variation in F/B ratio between sampling regions within the same patient and between patients in the same disease group compared with analyzing F and B data alone. The F/B ratio was independent of laser power drift, regions analyzed within a tissue and tissue orientation during analysis. Using this method, we identified a significant difference in collagen organization in airway tissue between COPD and non diseased. We have developed a robust optimization and calibration methodology that will allow direct comparison of data obtained at different times and from multiple microscopes, which is directly adaptable for use with other tissue types. We report a powerful new tool for advancing our understanding of pathological ECM remodeling that may uncover new therapeutic targets in the future.
细胞外基质 (ECM) 重塑有助于慢性阻塞性肺疾病 (COPD) 的发病变化，其既复杂又尚未被充分理解。在 COPD 气道中改变的 ECM 的胶原 I 具有二次谐波产生 (SHG) 特性。SHG 信号是相干的，沿与入射光平行的正向 (F)（主要为组织/成熟胶原纤维）和反向 (B)（主要为无序/不成熟胶原纤维）传播。F/B SHG 比用于确定有序胶原与无序胶原的比例，与单独分析 F 和 B 数据相比，同一患者内采样区域之间和同一疾病组内患者之间的 F/B 比值变化较小。F/B 比独立于激光功率漂移、组织内分析的区域以及组织在分析过程中的取向。使用该方法，我们在 COPD 和非病变气道组织中发现了胶原组织的显著差异。我们已经开发了一种强大的优化和校准方法，该方法将允许直接比较不同时间和来自多个显微镜的数据，并且可以直接适用于其他组织类型。我们报告了一种用于推进对病理性 ECM 重塑理解的强大新工具，该工具可能在未来揭示新的治疗靶点。
