After 0, 14 and 28 days of culture, constructs from each group (n=6) were harvested, lyophilized to obtain dry weight and digested with papain (125 μg/mL) at 60°C for 15 hours. dsDNA content was quantified using the Picogreen Assay (Life Technologies). Sulfated glycosaminoglycan (s-GAG) content was quantified spectrophotometrically using 1,9-dimethylmethylene blue (DMMB) dye (pH 3.0) with bovine chondroitin-4-sulfate (Sigma Aldrich) as a standard [43 (link)]. The hydroxyproline assay was used to determine total collagen content using a conversion factor of 1:7.46 to convert hydroxyproline to collagen [44 (link)]. All results were normalized to dry weight and reported as mean±SD.
Bovine chondroitin 4 sulfate
Manufactured by Merck Group
Bovine chondroitin-4-sulfate is a chemical compound derived from the cartilage of bovine (cattle) sources. It is a glycosaminoglycan composed of repeating disaccharide units of N-acetylgalactosamine and glucuronic acid. The primary function of this compound is to provide structural support and lubrication in cartilage and connective tissues.
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Lab products found in correlation
2 protocols using bovine chondroitin 4 sulfate
1
Quantifying Extracellular Matrix Components
2
Quantifying GAGs in Disc Tissue
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For GAGs level determination, total protein extract from disc tissue was measured using a 1,9-dimethyl methylene blue (DMMB) method with Bovine chondroitin 4-sulfate(Sigma-Aldrich) as a standard. 200 µl DMMB solution was added to the wells on a 96-well plate containing 40 µl of a sample. The DMMB solution was made by mixing 16 mg DMB, 3.04 g glycine, 1.6 g NaCl and 95 ml of 0.1 M Acetic Acid in 1 L of distilled water. The absorbance was measured at λ = 525nm on a SpectraMax M5 system (Molecular Devices, San Jose, CA, USA).
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