Table 2.
Traditional decellularized material characterization techniques
| Method | Components Identified | |
|---|---|---|
| Histology (Common stains include hematoxylin and eosin, Masson’s Trichrome, and Pentachrome) | Spatial location of remnant nucleic acid Tissue spatial organization | Spatial (Yes), Quantitative (No) |
| IHC/IF | Specific ECM proteins/structures of interest i.e. Collagen type, growth factors, immunogenic proteins |
Spatial (Yes), Quantitative (No) |
| Mass Spec | Quantify protein/biomolecule composition | |
| ToF-SIMS | Surface analytic compositional technique | Spatial (Yes), Quantitative (Yes) |
| Tandem MS | Bulk analytic compositional technique | Spatial (Yes), Quantitative (No) |
| Biopolymer Assays | Specific biopolymer identification | |
| Collagen (Hydroxyproline, Sircol™) | Mass fraction of collagen | Spatial (No), Quantitative (Yes) |
| GAG/Proteoglycan (Dimethylmethylene blue, Blyscan™) | Mass fraction of GAG | Spatial (No), Quantitative (Yes) |
| Elastin (Ninhydrin, Fastin™) | Mass fraction of Elastin | Spatial (No), Quantitative (Yes) |
| ELISA | Detection of soluble protein (growth factors) | Spatial (No), Quantitative (Yes) |
| Nucleic Acid (Hoescht based, Picogreen™, gel electrophoresis) | Rresidual DNA | Spatial (No), Quantitative (Yes) |
| Endotoxin (Limulus amebocyte lysate [LAL]) | Contaminating endotoxins | Spatial (No), Quantitative (Yes) |
| Mechanical Testing | ||
| Indentation | Modulus, Poisson ratio, permeability | Spatial (No), Quantitative (Yes) |
| Tensile | Stress-strain/Modulus | Spatial (No), Quantitative (Yes) |
| Thermal Denaturation | Crosslinking status | Spatial (No), Quantitative (Yes) |
| Fluorescence recovery after photobleaching (FRAP) | Small molecule diffusivity within decellularized construct | Spatial (No), Quantitative (Yes) |
| Laser Microscopy | Collagen, elastic fiber spatial organization | Spatial (Yes), Quantitative (No) |