Relation between micro- and nanostructure features and biological properties of the decellularized rat liver

Abstract

Organ decellularization is one of the promising technologies of regenerative medicine, which allows obtaining cell-free extracellular matrix (ECM), which provide preservation of the composition, architecture, vascular network and biological activity of the ECM. The method of decellularization opens up wide prospects for its practical application not only in the field of creating full-scale bioengineered structures, but also in the manufacture of vessels, microcarriers, hydrogels, and coatings. The main goal of our work was the investigation of structure and biological properties of lyophilized decellularized Wistar rat liver fragments (LDLFs), as well as we assessed the regenerative potential of the obtained ECM. We obtained decellularized liver of a Wistar rat, the vascular network and the main components of the ECM of tissue were preserved. H&E staining of histological sections confirmed the removal of cells. DNA content of ECM is equal to 0.7% of native tissue DNA content. Utilizing scanning probe nanotomogrphy method, we showed sinuous, rough topography and highly nanoporous structure of ECM, which provide high level of mouse 3T3 fibroblast and Hep-G2 cells biocompatibility. Obtained LDLF had a high regenerative potential, which we studied in an experimental model of a full-thickness rat skin wound healing: we observed the acceleration of wound healing by 2.2 times in comparison with the control.

Scaffolds Based on Silk Fibroin with Decellularized Rat Liver Microparticles: Investigation of the Structure, Biological Properties and Regenerative Potential for Skin Wound Healing

“…It makes sense to compare observed acceleration of skin regeneration with the results obtained previously for other scaffolds studied in the similar full-thickness rat skin woundhealing model, namely electrospun microfibrous scaffolds based on silk fibroin [36] and lyophilized decellularized Wistar rat liver fragments (LDLFs) [35].…”

“…This may also have been connected with the tissue-specific properties of the microparticles of decellularized tissue [44] or due to the biochemical composition of microparticles exposed on the film's surface. According to a previous study [35], the biochemical composition of the extracellular matrix after liver decellularization remained, including different types of collagens, which play a crucial role in the adhesion and proliferation of cells owing to RGD sequences in their structure.…”

“…A perfusion rate of 150 mL/h was used in all stages. The obtained decellularized liver was stored at 4 • C until use [35].…”

“…SEM analysis of the prepared samples was conducted using a Tescan Vega 3 (Tescan, Brno, Czech Republic) scanning electron microscope. Images were processed by Tescan VegaTC software version 4.2.17.0 (Tescan, Brno, Czech Republic) [35].…”

“…Samples for SPNT analysis were fixed, dehydrated, and embedded in epoxy embedding medium (Sigma-Aldrich, USA) using a protocol described in [35]. Consecutive ultramicrotomy sectioning of the samples was conducted using an ultrasonic diamond knife Ultrasonic 35 (Diatome AG, Nidau, Switzerland) with 3.0 mm blade width.…”