Raman spectroscopy of bone composition during healing of subcritical calvarial defects

Subcritical calvarial defects heal spontaneously and optical methods can study the healing without mechanically perturbing the bone. In this study, 1mm defects were created on the skulls (in vivo) of Sprague-Dawley rats (n = 14). After 7 (n = 7) and 14 days (n = 7) of healing, the subjects were sacrificed and additional defects were similarly created (control). Raman spectroscopy (785nm) was performed at the two time points and defect types. Spectra were quantified by the mineral/matrix ratio, carbonate/phosphate ratio and crystallinity. Mineral/matrix of in vivo defects is lower than that of controls by ~34% after 7 days and ~21% after 14 days. Carbonate/phosphate is 8% and 5% higher while crystallinity is 7% and 3% lower, respectively. Optical profiling shows that the surface roughness increases 1.2% from controls to in vivo after 7 days, then decreases 13% after 14 days. Overall, the results show maturation of mineral crystals during healing and agree with microscopic assessment.