Identification of tissue sections from decellularized liver scaffolds for repopulation experiments.

Area of Science:

• Regenerative Medicine
• Organ Engineering
• Tissue Engineering

Background:

• Biological organ engineering aims to create functional liver grafts through decellularization and repopulation.
• Current methods use healthy or diseased organs, but the impact of morphological changes on cell-scaffold interactions is unknown.
• This study addresses the need to identify liver scaffold sections with preserved microarchitecture for successful repopulation.

Purpose of the Study:

• To develop and validate a sequential morphological workflow for assessing decellularized liver scaffolds.
• To identify murine liver scaffold sections with well-preserved microarchitecture suitable for repopulation experiments.
• To evaluate the effectiveness of the workflow across native and pathologically altered liver scaffolds.

Main Methods:

• Decellularization of native and pathologically altered livers (steatosis, fibrosis, hyperplasia) using SDS and Triton X-100.
• A sequential morphological workflow involving macroscopic, microscopic, and histological/immunohistochemical assessments.
• Scaffold evaluation using decellularization scores at macroscopic, microscopic, and detailed histological/immunohistochemical levels.

Main Results:

• Decellularization was feasible across all experimental groups.
• A good macroscopic decellularization result was achieved in 11 of 33 scaffolds.
• The workflow identified 88 cell-free tissue sections, with 27 showing good histological and all groups yielding good immunohistochemical results.

Conclusions:

• Decellularization is achievable regardless of the liver's initial morphological state.
• The proposed sequential morphological workflow effectively identifies liver tissue sections with well-preserved microarchitecture.
• This method is crucial for selecting suitable scaffolds for subsequent repopulation in organ engineering.