Quantitative Collagen Biochemical Assay

Specimens from all four groups were removed following 22 days (n = 3 per group) and subjected to collagen biochemical assays (Biocolor life science assays, Carrickfergus, County Antrim, UK). Digested collagen samples were quantified similar to methods described previously [28 (link)]. Samples were digested with a solution of 0.5 M acetic acid (Sigma) and pepsin (1 mg/ml Pepsin (P7000), Sigma). Digestions were carried out for 16 hrs on a rocker (Orbitron Rotator; Boekel Scientific, Feasterville, PA) at 4°C. Collagen extracts were then assayed according to the in vitro tissue procedure provided by Sircol soluble collagen assay kit (Biocolor Ltd.). A multi-mode microplate reader was (Synergy HT, Biotek instrument, Inc, # 7091000) was set to an absorbance of 555 nm to obtain the collagen concentration.

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Rath S., Salinas M., Villegas A.G, & Ramaswamy S. (2015). Differentiation and Distribution of Marrow Stem Cells in Flex-Flow Environments Demonstrate Support of the Valvular Phenotype. PLoS ONE, 10(11), e0141802.

Publication 2015

Pepsin (P7000), Sircol soluble collagen assay kit Synergy HT

Acetic acid Assay Collagen Digestions Pepsin Tissue

Corresponding Organization : Florida International University

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Variable analysis

independent variables

Time (22 days)

dependent variables

Collagen concentration

control variables

Number of specimens per group (n = 3)

controls

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