Capturing effects of blood flow on the transplanted decellularized nephron with intravital microscopy

Abstract

Organ decellularization creates cell-free, collagen-based extracellular matrices that can be used as scaffolds for tissue engineering applications. This technique has recently gained much attention, yet adequate scaffold repopulation and implantation remain a challenge. Specifically, there still needs to be a greater understanding of scaffold responses post-transplantation and ways we can improve scaffold durability to withstand the in vivo environment. Recent studies have outlined vascular events that limit organ decellularization/recellularization scaffold viability for long-term transplantation. However, these insights have relied on in vitro/in vivo approaches that need enhanced spatial and temporal resolutions to investigate such issues at the microvascular level. This study uses intravital microscopy to gain instant feedback on their structure, function, and deformation dynamics. Thus, the objective of this study was to capture the effects of in vivo blood flow on the decellularized glomerulus, peritubular capillaries, and tubules after autologous and allogeneic orthotopic transplantation into rats. Large molecular weight dextran molecules labeled the vasculature. They revealed substantial degrees of translocation from glomerular and peritubular capillary tracks to the decellularized tubular epithelium and lumen as early as 12 h after transplantation, providing real-time evidence of the increases in microvascular permeability. Macromolecular extravasation persisted for a week, during which the decellularized microarchitecture was significantly and comparably compromised and thrombosed in both autologous and allogeneic approaches. These results indicate that in vivo multiphoton microscopy is a powerful approach for studying scaffold viability and identifying ways to promote scaffold longevity and vasculogenesis in bioartificial organs.

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

“…Also, as anticipated, histological assessments allowed the visualization of nuclei and the ECM components (primarily collagen) in native sections. In comparison, such assessments highlight the effective removal of cellular compartments and retention of ECM components and confirm successful decellularization ( Corridon et al, 2017 ; Garreta et al, 2017 ; Gilpin and Yang, 2017 ; Schmitt et al, 2017 ; Hillebrandt et al, 2019 ; Corridon, 2021 ; Polisetti et al, 2021 ; Wang et al, 2022a ; Shakeel and Corridon, 2022b ; Wang et al, 2022b ; Corridon, 2022 ; Neishabouri et al, 2022 ; Pantic et al, 2022 ; Zhang et al, 2022 ; Corridon, 2023b ; Khan et al, 2023 ).…”

A proposed model of xeno-keratoplasty using 3D printing and decellularization

“…Brightfield microscopy and hematoxylin-based DNA staining protocols are widely used for evaluating nuclear debris, and it is typically employed in conjunction with eosin to evaluate the fundamental ECM architecture of DCs ( Sc et al, 2011 ; Fu and Hollick, 2023 ; Vacalebre et al, 2023 ). Fluorescent nuclear stains such as DAPI ( Du and Wu, 2011 ; Gonzalez-Andrades et al, 2011 ; Shafiq et al, 2012 ), Hoechst ( Amano et al, 2008 ; Wu et al, 2009 ; Corridon, 2022 ; Corridon, 2023c ), and propidium iodide ( Shafiq et al, 2012 ) are also frequently utilized.…”