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Characterization of Extracellular Matrix Derived From Porcine Organs Decellularized Using Different Methods
Vignesh Dhandapani1,2, Pakindame Boabekoa1,2, Martin Borduas3
1Laboratoire de Bio-ingénierie et de Biophysique de l'Université de Sherbrooke, Department of Chemical and Biotechnological Engineering, Université de Sherbrooke, Québec, Canada.
Summary
This study characterizes extracellular matrices (ECMs) from decellularized porcine organs. Different methods yield ECMs with varying protein content and collagen structure, crucial for regenerative medicine applications.
Area of Science:
- Regenerative Medicine
- Biomaterials Science
- Tissue Engineering
Background:
- Regenerative medicine aims to create tissues and organs, potentially solving organ transplantation shortages.
- The extracellular matrix (ECM) is vital, providing a scaffold and biochemical signals for cell growth and differentiation.
- ECM composition includes fibrous proteins and proteoglycans, offering a dynamic microenvironment for cellular processes.
Purpose of the Study:
- To characterize extracellular matrices (ECMs) derived from porcine organs using four distinct decellularization techniques.
- To evaluate the impact of decellularization methods on ECM ultrastructure, composition, and residual cellular material.
- To establish a multimodal approach for assessing ECM quality for regenerative medicine.
Main Methods:
- Decellularization of porcine organs using four different methods.
- Histological analysis (Hematoxylin and eosin) to assess cellular removal and ECM integrity.
- Biochemical assays including DNA quantification, Bicinchoninic acid (BCA) assay for protein content.
- Mass spectrometry and proteomic analysis for detailed protein composition.
- Polarization microscopy to examine collagen fiber orientation.
Main Results:
- Histology confirmed the absence of nuclei and presence of glycosaminoglycans (GAGs) and collagen in ECMs.
- Native pancreas showed necrosis, impacting ECM quality, with reduced dsDNA content.
- Mass spectrometry confirmed significant differences in protein composition (up to 2700 proteins) based on decellularization technique.
- Residual double-stranded DNA (dsDNA) content in ECMs was lower than in native organs.
- BCA assay indicated varying protein content across different organs and techniques.
- Polarization microscopy revealed differences in collagen fiber orientation.
Conclusions:
- Decellularization techniques significantly influence the resulting ECM's protein composition and structural integrity.
- A multimodal characterization approach is essential for understanding ECM properties derived from different decellularization methods.
- Optimizing decellularization is key to balancing cellular component removal with the preservation of ECM ultrastructure and composition for regenerative applications.
