Raman investigation of in vivo radiation exposure on melanin in murine hair

Determining the effects of ionizing radiation from unintended exposure in a nuclear event requires the identification of relevant biomarkers and development of methods to retrospectively estimate the absorbed dose. Melanin, a biologically important natural pigment found in hair, shows promise as a biomarker to assess potential radiation exposure. We investigated Raman spectroscopy as a rapid and noninvasive technique to assess changes in melanin from the hair of C57BL/6 mice to gamma radiation between 0 and 4 Gy. Two excitation wavelengths (532 and 785 nm) were employed to probe the melanin response for changes with radiation exposure. Excitation wavelength-dependent variation in Raman features indicates resonance Raman effects, where a 785-nm excitation is more sensitive to the effects of gamma radiation. Melanin-specific Raman features were identified as potential biomarkers for gamma-radiation exposure and used to distinguish between irradiated and nonirradiated mice. Partial least square discriminant analysis models of exposure exhibited enhanced sensitivity to irradiation at 785 nm excitation and yielded a sensitivity of 88% and a specificity of 83%. Mice were classified with 100% sensitivity and specificity up to day 7 at a known time point. A decline in specificity and classification accuracy correlated with alterations in melanin's spectra after >7 days following irradiation. Regression models of the Raman spectrum determined the exposed dose with a precision of <1 Gy at a known exposure time point. This noninvasive approach offers promising applications in radiation biodosimetry and medical monitoring, providing retrospective detection of gamma-radiation exposure at clinically relevant doses.