Abstract
Tissue decellularization has rapidly developed to be a practical approach in tissue engineering research; biological tissue is cleared of cells resulting in a protein-rich husk as a natural scaffold for growing transplanted cells as a donor organ therapy. Minimally processed, acellular extracellular matrix reproduces natural interactions with cells in vitro and for tissue engineering applications in animal models. There are many decellularization techniques that achieve preservation of molecular profile (proteins and sugars), microstructure features such as organization of ECM layers (interstitial matrix and basement membrane) and organ level macrofeatures (vasculature and tissue compartments). While structural and molecular cues receive attention, mechanical and material properties of decellularized tissues are not often discussed. The effects of decellularization on an organ depend on the tissue properties, clearing mechanism, chemical interactions, solubility, temperature, and treatment duration. Physical characterization by a few labs including work from the authors provides evidence that decellularization protocols should be tailored to specific research questions. Physical characterization beyond histology and immunohistochemistry of the decellularized matrix (dECM) extends evaluation of retained functional features of the original tissue. We direct our attention to current technologies that can be employed for structure function analysis of dECM using underutilized tools such as atomic force microscopy (AFM), cryogenic electron microscopy (cryo-EM), dynamic mechanical analysis (DMA), Fourier-transform infrared spectroscopy (FTIR), mass spectrometry, and rheometry. Structural imaging and mechanical functional testing combined with high-throughput molecular analyses opens a new approach for a deeper appreciation of how cellular behavior is influenced by the isolated microenvironment (specifically dECM). Additionally, the impact of these features with different decellularization techniques and generation of synthetic material scaffolds with desired attributes are informed. Ultimately, this mechanical profiling provides a new dimension to our understanding of decellularized matrix and its role in new applications.
Highlights
Tissue decellularization has rapidly developed to be a practical approach in tissue engineering research; biological tissue is cleared of cells resulting in a protein-rich husk as a natural scaffold for growing transplanted cells as a donor organ therapy
We direct our attention to current technologies that can be employed for structure function analysis of decellularized matrix (dECM) using underutilized tools such as atomic force microscopy (AFM), cryogenic electron microscopy, dynamic mechanical analysis (DMA), Fourier-transform infrared spectroscopy (FTIR), mass spectrometry, and rheometry
In our review of decellularization papers, we found that scanning electron microscopy (SEM) was the most frequently employed technique for dECM structural analysis
Summary
Extracellular matrix (ECM) is a complex structure which physically supports cells in virtually every tissue in the body. ECM remodeling involves both biochemical and mechanical alterations maintained in biophysical feedback loops These feedback loops are sustained by biochemical and mechanical signaling between cell and cell-microenvironment. The importance of structural and mechanical cues provided by the ECM has direct implications for the usefulness of isolated biological scaffolds and has been comprehensively reviewed in recent times. Relevant for cell-substrate interactions is the review on ECM physical cues, “Extracellular matrix elasticity and topography: material-based cues that affect cell function via conserved mechanisms” by Janson and Putnam [17]. Including, but not limited to, western blot methods and immunohistochemistry These two common techniques, are not the most suitable for characterizing critical structural and mechanical ECM cues
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