In Situ Monitoring of Chondrocyte Response to Bioactive Scaffolds Using Raman Spectroscopy

Twenty-first century challenges for biomaterials

“…Raman spectral signatures can be obtained with a high signal-to-noise ratio on living cells with 2 min exposure to 115 mW of 785 nm laser light without damage to the cells (Notingher et al 2002. A series of investigations showed that the bio-Raman method makes it possible to monitor the viability, cell cycle, metabolism, mitosis, differentiation, de-differentiation, mineralization and onset of death of single cells and cell assemblages (organoids) in real time without damage to the cells (Notingher et al 2002(Notingher et al ,b, 2004a(Notingher et al -e, 2005Owen et al 2004Owen et al , 2006aVerrier et al 2004b;Jones et al 2005;Chan et al 2006;Notingher & Hench 2006;Swain & Stevens 2007;Jell et al 2008;Pyrgiotakis et al 2008;Swain et al 2008a,b;Gentleman et al 2009). Single cells or three-dimensional assemblages of cells living in a specially designed microscope chamber can survive for long periods of time (several days) while being interrogated as to their state of health without alteration or damage to the cells (Notingher et al 2002.…”

“…An LDA model prepared from the data showed high cross-validation sensitivity (100%) and high specificity (95%) for discriminating the MG63 cells from the primary cells, with 96 per cent of the cells being correctly classified either as tumour-derived or non-tumourderived cells. A similar analytical method has been used to monitor cell cycle dynamics (Swain et al 2008b) and to analyse the molecular changes of stem cells undergoing differentiation towards a specific cell phenotype (Notingher et al 2004a,b,d;Jones et al 2005;Chan et al 2006;Jell et al 2008;Swain et al 2008a). …”

“…Differences between the biological actions of cytotoxic agents (Triton X-100) versus genotoxic agents (Etoposide, VP16) can be detected by use of bio-Raman spectroscopy. This is achieved by determining the mechanism of cell death by either disruption of the cell membrane (cytotoxin) or breaking of DNA (genotoxin; Notingher et al 2003aNotingher et al , 2004cNotingher et al ,e, 2005Owen et al 2004Owen et al , 2006aVerrier et al 2004b;Notingher & Hench 2006;Swain & Stevens 2007). This bio-Raman methodology was used to achieve an important milestone in our DARPA-funded research programme; i.e.…”

“…The bio-Raman system offers the potential of monitoring and maintaining cell phenotype by in situ spectroscopic analysis of the cells Jones et al 2005;Jell et al 2008;Swain et al 2008a;Gentleman et al 2009). Notingher et al show that two statistical-based spectroscopic data management programs, called principal component analysis and linear discriminant analysis (LDA), are powerful tools for distinguishing cell phenotypes in real time.…”

1984 : on monitoring cell fate in three-dimensional polymeric scaffolds for tissue engineering applications

“…Spectral information retrieved by scanning specimens over a certain area, optionally confocal, can be computed and displayed as (3D) cluster images [196]. Raman spectroscopy and Raman spectroscopy based imaging has been successfully applied in several tissue engineering applications for example by monitoring chondrocyte behavior on bioactive scaffolds [197]. Boyd et al have utilized a bench-top macro-Raman spectrometer with high-throughput screening capability to examine spectral differences between well characterized cell lines (two types of osteosarcoma cells, human dermal fibroblasts and human embryonic lung epithelial cells) and the effects of cellular death on the retrieved spectra [198].…”