Examining lysyl oxidase-like modulation of collagen architecture in 3D spheroid models of idiopathic pulmonary fibrosis via second-harmonic generation microscopy.
Examining lysyl oxidase-like modulation of collagen architecture in 3D spheroid models of idiopathic pulmonary fibrosis via second-harmonic generation microscopy.
复制标题
用二次谐波产生显微镜检测特发性肺纤维化3D球体模型中赖氨酰氧化酶样胶原结构的调节。
DOI：
10.1117/1.jbo.26.6.066501
复制
发表时间：
2021-06
影响因子：
3.5
通讯作者：
Campagnola PJ
中科院分区：
文献类型：
--
作者：
James DS;Brereton CJ;Davies DE;Jones MG;Campagnola PJ
重要性：特发性肺纤维化（IPF）患者预后不良，诊断后寿命短，因为有效的治疗选择有限。尽管基质硬化是该疾病的标志，但仍然缺乏关于该疾病中潜在胶原改变的知识。具体而言，虽然涉及胶原交联增加，但尚未探索对胶原大分子/超分子变化的影响。目的：我们试图确定二次谐波发生（SHG）显微镜是否可以表征在不同交联调节条件（促进和抑制）下生长的IPF 3D球体模型中胶原结构的差异。方法：我们使用基于纤维形态、相对SHG亮度和通过SHG偏振分析的大分子/超分子结构的SHG度量来比较IPF球状体的结构。结果：球体纤维形态比较显示，对照组纤维最长、最直、最粗。具有交联增强和抑制的球状体分别具有最高和最低的SHG转换效率，这与所得到的谐波团密度一致。SHG偏振分析表明，肽的螺距角，胶原蛋白分子的排列，和整体手性交联调制后改变，也与减少组织相对于对照组一致。结论：虽然没有单一的SHG签名与交联相关，但我们表明，这里使用的一套指标在描绘胶原蛋白结构尺寸尺度的变化方面是有效的。结果在很大程度上反映了人体组织的结果，并证明3D球体模型和SHG分析的组合是假设检验IPF中操作性细胞和分子因子作用的有力方法。
Significance: Idiopathic pulmonary fibrosis (IPF) patients have a poor prognosis with short lifespan following diagnosis as there are limited effective treatment options. Despite matrix stiffening being the hallmark of the disease there remains a lack of knowledge surrounding the underlying collagen alterations in the disease. Specifically, while increased collagen crosslinking has been implicated, the resulting effects on collagen macro/supramolecular changes have not been explored. Aim: We sought to determine if second-harmonic generation (SHG) microscopy could characterize differences in the collagen architecture in 3D spheroid models of IPF grown under different crosslinking modulation conditions (promotion and inhibition). Approach: We used SHG metrics based on the fiber morphology, relative SHG brightness, and macro/supramolecular structure by SHG polarization analyses to compare the structure of the IPF spheroids. Results: Comparison of the fiber morphology of the spheroids showed that the control group had the longest, straightest, and thickest fibers. The spheroids with crosslink enhancement and inhibition had the highest and lowest SHG conversion efficiencies, respectively, consistent with the resulting harmonophore density. SHG polarization analyses showed that the peptide pitch angle, alignment of collagen molecules, and overall chirality were altered upon crosslink modulation and were also consistent with reduced organization relative to the control group. Conclusions: While no single SHG signature is associated with crosslinking, we show that the suite of metrics used here is effective in delineating alterations across the collagen architecture sizescales. The results largely mirror those of human tissues and demonstrate that the combination of 3D spheroid models and SHG analysis is a powerful approach for hypothesis testing the roles of operative cellular and molecular factors in IPF.