Both Fourier Transform Infrared (FTIR) and Raman spectroscopy have been applied to thein vitrocharacterisation of biomaterials, mainly surface reactions leading to the formation of a biologically active hydroxycarbonate apatite (HCA) layer on the sample surface when immersed in simulated body fluids (SBF). The HCA layer indicates the degree of bioactivity of the sample, because it leads to a strong bond between the biomaterial and living tissue. Reflection measurements using FTIR allow quick, non-destructive detection of the HCA layer for solid and powder samples. Due to the low Raman scattering efficiency and low absorption of water in the visible-near infrared region, Raman micro-spectroscopy was successfully used for thein situcharacterisation of 20 and 40µm diameter 45S5 Bioglass®fibres. Thein situcapabilities of the Raman micro-spectrometer have also been extended to the characterisation of living cells attached on bioinert silica and bioactive 45S5 Bioglass®and 58S substrates. Using a high power 785 nm laser, living cells in physiological conditions can be real-time sampled over long periods of time without inducing cell damage and with good signal strength. Cell death can be monitored because it proved to induce strong changes in the Raman signature in the spectral regions 1000–1150 cm–1and 1550–1650 cm–1.
Bioactive glass fibres can be used as tissue engineering scaffolds. In this investigation, the bioactive response of 45S5 glass fibres was assessed in simulated body fluid (SBF). Preliminary attachment of osteoblasts to the fibre surface was assessed, as were the fibre tensile strength and fracture toughness. Fourier transform infrared spectroscopy (FTIR) analysis revealed that hydroxyapatite (HA) was formed on the fibres' surface after 2-4 days in SBF. Raman micro-spectroscopic analysis was used to monitor development of the HA layer during immersion. A correlation was found between increase in intensity of the PO4(3-) peak near 964cm(-1) and appearance of crystalline HA (P-O bending peaks) using FTIR. Such results are encouraging for in situ bioactivity monitoring, as Raman spectra are insensitive to the presence of water, unlike FTIR. Average tensile strength of 45S5 fibres (79 microm diameter) was 340+/-140 MPa. Fracture toughness, determined using fracture surface analysis, was 0.7+/-0.1 MPa m1/2. Confocal microscopy revealed osteoblasts attached and spread along the fibres after 15-90 min culture. Scanning electron microscopy analysis showed that cells with filopodia and dorsal ruffles remained attached after 14 days in culture. These results are encouraging, as cell adhesion is an important first step prior to proliferation and differentiation.
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