Tissue decellularization has emerged as a technique to provide an acellular, non-immunogenic scaffold that preserves the morphological features of native tissue. To study the possible effects of decellularization, investigating the mechanical behavior and the protein composition is crucial. In this study, we performed extension-inflation tests on native and decellularized porcine vena cava and investigated their microstructure using multiphoton microscopy. The mechanical behavior of both groups showed typical pressure-stretch curves of vascular structures with viscoelastic and nonlinear features. Importantly, no significant differences were found at inflation of 10, 20 and 30 mmHg, although some variability was observed in the decellularized scaffolds. When analyzing the results of the vessel wall multiphoton microscopy investigations, it was found that collagen fibers were packed in tortuous bundles in the media, but scattered in the adventitia. The fibers were oriented around 72° from the circumferential direction for both groups and at the same time equally distributed out-of-plane. Moreover, the collagen fibers diameter for media and adventitia was around 4 μm. Tortuosity and straightness were the same in the adventitia; however, the situation was different in the media, where the fibers in native samples were straighter than in decellularized scaffolds. Our findings show the potential of our protocol to obtain venous scaffolds that could be used for vascular reconstruction, as their mechanical properties are largely comparable to those of their native counterparts. The detailed analysis of the microstructure also represents a first step towards better understanding the physiology of the vessels and replicating these conditions in silico. STATEMENT OF SIGNIFICANCE: Tissue engineering provides a scaffold as substrate for in vitro cells seeding. Decellularization completely removes immunogenic cellular components, preserving the organ ultrastructure. Consequently, decellularized scaffolds provide a natural microenvironment for cell repopulation and facilitate functional recovery in vitro. We have comprehensively characterized the decellularized porcine vena cava by comparing its mechanical properties and microstructural characteristics with its native counterpart. Extension-inflation testing is considered a method to mimic stresses and stretches in vivo. Since no significant differences were found between native and decellularized tissue, these scaffolds show some potential. Moreover, this study was expanded to include microstructural characterization of collagen fibers using multi-photon microscopy, making it the first of its kind dedicated to biomechanical and microstructural evaluation of decellularized veins.

Biomedical Center Faculty of Medicine in Pilsen Charles University Czech Republic

Department of Biophysics Faculty of Medicine in Pilsen Charles University Czech Republic

Department of Structural Engineering NTNU Trondheim Norway

Department of Surgery Faculty of Medicine in Pilsen Charles University Czech Republic

Field of Excellence BioHealth University of Graz Austria

Institute of Biomechanics Graz University of Technology Austria

Institute of Molecular Biosciences University of Graz Austria

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