Detection of early osteogenic commitment in primary cells using Raman spectroscopy

The Analyst
Stephanie J SmithSumeet Mahajan

Abstract

Major challenges in the development of novel implant surfaces for artificial joints include osteoblast heterogeneity and the lack of a simple and sensitive in vitro assay to measure early osteogenic responses. Raman spectroscopy is a label-free, non-invasive and non-destructive vibrational fingerprinting optical technique that is increasingly being applied to detect biochemical changes in cells. In this study Raman spectroscopy has been used to obtain bone cell-specific spectral signatures and to identify any changes therein during osteoblast commitment and differentiation of primary cells in culture. Murine calvarial osteoblasts (COBs) were extracted and cultured and studied by Raman spectroscopy over a 14 day culture period. Distinct osteogenic Raman spectra were identified after 3 days of culture with strong bands detected for mineral: phosphate ν3 (1030 cm-1) and B-type carbonate (1072 cm-1), DNA (782 cm-1) and collagen matrix (CH2 deformation at 1450 cm-1) and weaker phosphate bands (948 and 970 cm-1). Early changes were detected by Raman spectroscopy compared to a standard enzymatic alkaline phosphatase (ALP) assay and gene expression analyses over this period. Proliferation of COBs was confirmed by fluorescence intensity...Continue Reading

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Citations

Jul 4, 2019·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Alice GoringClaire E Clarkin
Apr 5, 2019·Scientific Reports·Catarina Costa MouraSumeet Mahajan
Jun 13, 2021·Laboratory Investigation; a Journal of Technical Methods and Pathology·Pascaline BouzyNicholas Stone
Nov 18, 2021·The Analyst·Mustafa UnalJeffry S Nyman

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