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From Tissue to Insight: Advanced Tissue Processing and Spatial Imaging in Translational Lung Transplantation

Wang, Qi LU (2026) In Lund University, Faculty of Medicine Doctoral Dissertation Series
Abstract
Acute lung injury (ALI) and transplantation-related graft dysfunction remain major clinical and scientific challenges. In conditions relevant to donor lung assessment and transplantation, injury can affect multiple tissue compartments, including the alveolar epithelium, vascular endothelium, immune microenvironment, and extracellular matrix (ECM). These alterations can compromise the alveolar–capillary barrier, impair gas exchange and lung mechanics, and reduce donor lung quality or graft function. Understanding these processes requires investigation of how structural, cellular, and molecular changes are spatially distributed within lung tissue.

This thesis investigated tissue-level alterations in translational porcine models of... (More)
Acute lung injury (ALI) and transplantation-related graft dysfunction remain major clinical and scientific challenges. In conditions relevant to donor lung assessment and transplantation, injury can affect multiple tissue compartments, including the alveolar epithelium, vascular endothelium, immune microenvironment, and extracellular matrix (ECM). These alterations can compromise the alveolar–capillary barrier, impair gas exchange and lung mechanics, and reduce donor lung quality or graft function. Understanding these processes requires investigation of how structural, cellular, and molecular changes are spatially distributed within lung tissue.

This thesis investigated tissue-level alterations in translational porcine models of aspiration-induced donor lung injury and complement-mediated graft injury, with a focus on alveolar architecture, barrier integrity, immune activation, and ECM organisation. Tissue-processing and sectioning methods were first evaluated to support histological and fluorescence-based analyses. Isopropanol provided histological quality, sectioning performance, and fluorescence-based measurements comparable to those obtained with xylene. Agarose embedding combined with automated vibratome sectioning enabled the preparation of structurally preserved lung sections for high-resolution multiplex fluorescence imaging.

Quantitative fluorescence imaging showed that aspiration-induced injury increased alveolar wall thickness and reduced alveolar circularity. These features were integrated into a morphological quotient (MQ) that captured injury-related changes in alveolar architecture. Across the aspiration-induced and complement-mediated injury models, spatial analysis revealed alterations involving the alveolar epithelium, epithelial junctions, vascular endothelium, innate immune response, ECM, and contractile structures, with both shared and model-specific patterns. These tissue changes occurred alongside functional deterioration and histological injury in both models.

Overall, the workflows evaluated and applied in this thesis enabled quantitative characterisation of multicompartment lung tissue injury. Spatial tissue analysis provided information complementary to conventional histology and physiological assessment and distinguished tissue responses shared by or
specific to the two injury models. This work provides a basis for further validation of spatial imaging approaches in mechanistic studies, donor lung assessment, and evaluation of therapeutic responses in lung transplantation. (Less)
Please use this url to cite or link to this publication:
author
supervisor
opponent
  • docent Oltean, Mihai, Göteborg University
organization
publishing date
type
Thesis
publication status
published
subject
keywords
lung transplantation, Donor lung evaluation, Ex vivo lung perfusion, Large animal model, Tissue processing, Multiplex immunofluorescence, Quantitative spatial analysis
in
Lund University, Faculty of Medicine Doctoral Dissertation Series
issue
2026:108
pages
93 pages
publisher
Lund University, Faculty of Medicine
defense location
Belfragesalen, BMC D15, Klinikgatan 32 i Lund
defense date
2026-09-03 08:30:00
ISSN
1652-8220
ISBN
978-91-8021-906-8
language
English
LU publication?
yes
id
9012507e-89a5-4063-8ec5-51b6978b58ab
date added to LUP
2026-08-07 11:50:41
date last changed
2026-08-14 13:01:16
@phdthesis{9012507e-89a5-4063-8ec5-51b6978b58ab,
  abstract     = {{Acute lung injury (ALI) and transplantation-related graft dysfunction remain major clinical and scientific challenges. In conditions relevant to donor lung assessment and transplantation, injury can affect multiple tissue compartments, including the alveolar epithelium, vascular endothelium, immune microenvironment, and extracellular matrix (ECM). These alterations can compromise the alveolar–capillary barrier, impair gas exchange and lung mechanics, and reduce donor lung quality or graft function. Understanding these processes requires investigation of how structural, cellular, and molecular changes are spatially distributed within lung tissue.<br/><br/>This thesis investigated tissue-level alterations in translational porcine models of aspiration-induced donor lung injury and complement-mediated graft injury, with a focus on alveolar architecture, barrier integrity, immune activation, and ECM organisation. Tissue-processing and sectioning methods were first evaluated to support histological and fluorescence-based analyses. Isopropanol provided histological quality, sectioning performance, and fluorescence-based measurements comparable to those obtained with xylene. Agarose embedding combined with automated vibratome sectioning enabled the preparation of structurally preserved lung sections for high-resolution multiplex fluorescence imaging.<br/><br/>Quantitative fluorescence imaging showed that aspiration-induced injury increased alveolar wall thickness and reduced alveolar circularity. These features were integrated into a morphological quotient (MQ) that captured injury-related changes in alveolar architecture. Across the aspiration-induced and complement-mediated injury models, spatial analysis revealed alterations involving the alveolar epithelium, epithelial junctions, vascular endothelium, innate immune response, ECM, and contractile structures, with both shared and model-specific patterns. These tissue changes occurred alongside functional deterioration and histological injury in both models.<br/><br/>Overall, the workflows evaluated and applied in this thesis enabled quantitative characterisation of multicompartment lung tissue injury. Spatial tissue analysis provided information complementary to conventional histology and physiological assessment and distinguished tissue responses shared by or <br/>specific to the two injury models. This work provides a basis for further validation of spatial imaging approaches in mechanistic studies, donor lung assessment, and evaluation of therapeutic responses in lung transplantation.}},
  author       = {{Wang, Qi}},
  isbn         = {{978-91-8021-906-8}},
  issn         = {{1652-8220}},
  keywords     = {{lung transplantation; Donor lung evaluation; Ex vivo lung perfusion; Large animal model; Tissue processing; Multiplex immunofluorescence; Quantitative spatial analysis}},
  language     = {{eng}},
  number       = {{2026:108}},
  publisher    = {{Lund University, Faculty of Medicine}},
  school       = {{Lund University}},
  series       = {{Lund University, Faculty of Medicine Doctoral Dissertation Series}},
  title        = {{From Tissue to Insight: Advanced Tissue Processing and Spatial Imaging in Translational Lung Transplantation}},
  url          = {{https://lup.lub.lu.se/search/files/257411309/Kappa_Qi_Wang_26-07-28.pdf}},
  year         = {{2026}},
}