Elastin and glycosaminoglycan were quantified as previously described26 (link).
The elastin content of native and decellularized tissues was quantified using the FASTIN elastin assay (Biocolor) according to the manufacturer’s instructions. Briefly, the samples were homogenized and elastin was solubilized in 0.25 M oxalic acid. Two consecutive incubations were performed at 95 °C to ensure complete extraction of elastin. Extracts were incubated with 5,10,15,20-tetraphenyl-21H,23H-porphine tetrasulfonate (TPPS) dye and absorbance was determined at 555 nm spectrophotometrically (Tecan Infinity). Elastin concentrations from a standard curve were used to calculate the elastin content of the tissue.
The Glycosaminoglycan (sGAG) content of native and decellularized tissues was quantified using the Blyscan GAG Assay Kit (Biocolor). Briefly, the tissues were digested with papain at 65 °C for 18 h and aliquots of each sample were mixed with 1,9-dimethyl-methylene blue dye and reagents from the GAG assay kit. The absorbance at 656 nm was measured spectrophotometrically (Tecan Infinity) and compared to standards made from bovine tracheal chondroitin-4-sulfate to determine the sGAG content.
The elastin content of native and decellularized tissues was quantified using the FASTIN elastin assay (Biocolor) according to the manufacturer’s instructions. Briefly, the samples were homogenized and elastin was solubilized in 0.25 M oxalic acid. Two consecutive incubations were performed at 95 °C to ensure complete extraction of elastin. Extracts were incubated with 5,10,15,20-tetraphenyl-21H,23H-porphine tetrasulfonate (TPPS) dye and absorbance was determined at 555 nm spectrophotometrically (Tecan Infinity). Elastin concentrations from a standard curve were used to calculate the elastin content of the tissue.
The Glycosaminoglycan (sGAG) content of native and decellularized tissues was quantified using the Blyscan GAG Assay Kit (Biocolor). Briefly, the tissues were digested with papain at 65 °C for 18 h and aliquots of each sample were mixed with 1,9-dimethyl-methylene blue dye and reagents from the GAG assay kit. The absorbance at 656 nm was measured spectrophotometrically (Tecan Infinity) and compared to standards made from bovine tracheal chondroitin-4-sulfate to determine the sGAG content.
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