Ceramic materials lead to underestimated DNA quantifications: a method for reliable measurements

European Cells & Materials
E PiccininiI Martin

Abstract

In the context of investigating cell-material interactions or of material-guided generation of tissues, DNA quantification represents an elective method to precisely assess the number of cells attached or embedded within different substrates. Nonetheless, nucleic acids are known to electrostatically bind to ceramics, a class of materials commonly employed in orthopaedic implants and bone tissue engineering scaffolds. This phenomenon is expected to lead to a relevant underestimation of the DNA amount, resulting in erroneous experimental readouts. The present work aims at *lpar;i) investigating the effects of DNA-ceramic bond occurrence on DNA quantification, and (ii) developing a method to reliably extract and accurately quantify DNA in ceramic-containing specimens. A cell-free model was adopted to study DNA-ceramic binding, highlighting an evident DNA loss (up to 90%) over a wide range of DNA/ceramic ratios (w/w). A phosphate buffer-based (800 mM) enzymatic extraction protocol was developed and its efficacy in terms of reliable DNA extraction and measurement was confirmed with commonly used fluorometric assays, for various ceramic substrates. The proposed buffered DNA extraction technique was validated in a cell-based experimen...Continue Reading

Citations

Feb 18, 2015·Journal of Cellular and Molecular Medicine·Benjamin E PippengerIvan Martin
Dec 12, 2013·Journal of Biomaterials Applications·Arina T BuizerRoel Kuijer
Apr 23, 2021·Xenotransplantation·Joseph Sushil RaoChristopher Burlak

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