Histological Analysis of Lung Tissue

Lung tissues were collected, fixed with 10% neutral buffered formalin (48 h) and embedded in paraffin fixation. Then, 4-μm thick sections (3 sections per animal) were cut and stained with hematoxylin and eosin (HE), periodic acid–Schiff (PAS), and Masson trichrome. All pictures were captured using a Nikon microscope (Tokyo, Japan). Peribronchial inflammation score (grades 0-4) was evaluated in a blind-way (24 (link)). Goblet cell hyperplasia (grades 0-4) was determined using the method described by Padrid et al. (25 (link)). Image-Pro Plus software (Version X; Adobe, San Jose, CA) was used to quantify the areas occupied by collagen (blue, Masson trichrome staining), which were subsequently divided by the total area examined (as the percentage of collagen fibers) (8 (link)). At least 6 bronchioles were counted in each slide, and then, the mean inflammation score, goblet cell hyperplasia score, and percentage of collagen fibers were calculated for each mouse.

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Xu X., Wang Y., Luo X., Gao X., Gu W., Ma Y., Xu L., Yu M., Liu X., Liu J., Wang X., Zheng T., Mao C, & Dong L. (2023). A non-invasive strategy for suppressing asthmatic airway inflammation and remodeling: Inhalation of nebulized hypoxic hUCMSC-derived extracellular vesicles. Frontiers in Immunology, 14, 1150971.

Publication 2023

Image-Pro Plus software

Animal Blind Bronchioles Collagen Embedded paraffin Eosin Formalin Goblet cell Hyperplasia Inflammation Lung Microscope Mouse Periodic acid Tissues

Corresponding Organization : Affiliated Hospital of Jiangsu University

Other organizations : Xuzhou Medical College, Second People’s Hospital of Huai’an, Jintan People's Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Peribronchial inflammation score (grades 0-4)
Goblet cell hyperplasia (grades 0-4)
Percentage of collagen fibers

control variables

Lung tissues were collected, fixed with 10% neutral buffered formalin (48 h) and embedded in paraffin fixation
4-μm thick sections (3 sections per animal) were cut and stained with hematoxylin and eosin (HE), periodic acid–Schiff (PAS), and Masson trichrome
All pictures were captured using a Nikon microscope (Tokyo, Japan)
At least 6 bronchioles were counted in each slide, and then, the mean inflammation score, goblet cell hyperplasia score, and percentage of collagen fibers were calculated for each mouse

controls

None explicitly mentioned

Annotations

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