http://www.bio.umass.edu/biology/kunkel/pub/lobster/PDFs/CrystalChemistry/Crane_OctaCaPO4-Bone.pdf

Raman spectroscopic evidence for octacalcium phosphate and other transient mineral species deposited during intramembranous mineralization.

Infrared (IR) and Raman spectroscopy are emerging biophotonic tools to recognize various diseases. The current review gives an overview of the experimental techniques, data-classification algorithms and applications to assess soft tissues, hard tissues and body fluids. The methodology section presents the principles to combine vibrational spectroscopy with microscopy, lateral information and fiber-optic probes. A crucial step is the classification of spectral data by a variety of algorithms. We discuss unsupervised algorithms such as cluster analysis or principal component analysis and supervised algorithms such as linear discriminant analysis, soft independent modeling of class analogies, artificial neural networks support vector machines, Bayesian classification, partial least-squares regression and ensemble methods. The selected topics include tumors of epithelial tissue, brain tumors, prion diseases, bone diseases, atherosclerosis, kidney stones and gallstones, skin tumors, diabetes and osteoarthritis.

Disease recognition by infrared and Raman spectroscopy.

In addition to mechanical functions, bones have an essential role in metabolic activity as mineral reservoirs that are able to absorb and release ions. Bioapatite, considered the major component in the mineralized part of mammalian bones, is a calcium phosphate mineral with a structure that closely resembles hydroxyapatite (HA, Ca10[PO4]6[OH]2) with variable chemical substitutions. It is important to note that it continues to be chemically active long after it has been initially deposited. Detailed understanding of changes in the mineral phase as HA matures is essential for understanding how normal bone achieves its remarkable mechanical performance, how it is altered in disease, as well as the effects of therapeutic interventions. A model system for investigation of the in vivo maturation of HA is available, namely, the in vitro conversion of amorphous calcium phosphate (ACP) to HA in a supersaturated solution of calcium and phosphate ions. In the present study, this system was employed to correlate with the changes in chemistry and poorly crystalline HAP crystal size, shape, and habit. The results of the X-ray diffraction as well as Raman analyses showed that as the crystallites mature in the 002 and 310 directions both the full width at half-height and wavelength at maximum of the Raman peaks change as a function of reaction extent and crystallite maturation, size, and shape. Moreover, such analyses can be performed in intact bone specimens through Raman microspectroscopic and imaging analyses with a spatial resolution of 0.6–1 μ, by far superior to the one offered by other microspectroscopic techniques, thus potentially yielding important new information on the organization and mineral quality of normal and fragile bone.

Complementary Information on In Vitro Conversion of Amorphous (Precursor) Calcium Phosphate to Hydroxyapatite from Raman Microspectroscopy and Wide-Angle X-Ray Scattering

Raman spectroscopy provides the unique opportunity to nondestructively analyze chemical concentrations in individual cells on the submicrometer length scale without the need for optical labels. This enables the rapid assessment of cellular biochemistry inside living cells, and it allows for their continued analysis. Here, we review recent developments in the analysis of single cells, subcellular compartments, and chemical imaging based on Raman spectroscopy. Spontaneous Raman spectroscopy provides for the full spectral assessment of cellular biochemistry, while coherent Raman techniques, such as coherent anti-Stokes Raman scattering is primarily used as an imaging tool comparable to confocal fluorescence microscopy. These techniques are complemented by surface-enhanced Raman spectroscopy, which provides higher sensitivity and local specificity, and also extends the techniques to chemical indicators, i.e. pH sensing. We review the strengths and weaknesses of each technique, demonstrate some of their applications and discuss their potential for future research in cell biology and biomedicine.

Raman spectroscopy and microscopy of individual cells and cellular components

On octacalcium phosphate

Craniosynostosis is a severe craniofacial disease where one or more sutures, the fibrous tissue that lies between the cranial bones, fuses prematurely. Some craniosynostosis syndromes are known to be caused by mutations in fibroblast growth factor (FGF) receptors. Mutated FGF receptors are thought to cause constitutive signaling. In this study, heparin acrylic beads released fibroblast growth factor 2 (FGF2) to mimic constitutive signaling by mutated receptors, delivering FGF2 in addition to already existing normal tissue amounts. Fetal day 18.5 mouse sutures were treated with FGF2-soaked beads and cultured in serum free media for 48 h. We have shown previously that this treatment leads to fusion and increased Msx2 expression, but here we use near-infrared Raman imaging to simultaneously examine the mineral components and matrix components of cranial tissue while providing light microscopic spatial information. FGF2-treated mouse sutures show increased v1 phosphate and v1 carbonate bandwidths, indicating a slightly chemically modified mineral being rapidly deposited. In addition, FGF2-treated mouse sutures show a marked increase in mineral-to-matrix ratios compared to control mouse sutures, typical of increased mineralization.

https://www.spiedigitallibrary.org/journals/Journal-of-Biomedical-Optics/volume-10/issue-03/031119/Raman-imaging-demonstrates-FGF2-induced-craniosynostosis-in-mouse-calvaria/10.1117/1.1908057.pdf

Raman imaging demonstrates FGF2-induced craniosynostosis in mouse calvaria.

Raman spectral imaging provides the means to study spatially localized response to controlled external perturbations to tissue specimens with light microscopy resolution. We discuss use of heparin-acrylic microbeads soaked with the protein fibroblast growth factor-2 (FGF2). Microbeads containing FGF2 are placed in murine calvarial tissue and stimulate abnormally rapid mineralization. The tissue response simulated the effects of craniosynostosis, a birth defect occurring in 1 in 2400 live births. We describe Raman imaging measurements of the spatial distribution of apatitic mineral and matrix (predominantly type I collagen) from normal murine calvarial tissue and murine calvarial tissue modeling craniosynostosis. We also discuss spectroscopic evaluation of the state of the mineral induced by the FGF2 beads.

Study of localization of response to fibroblast growth factor-2 in murine calvaria using Raman spectroscopic imaging

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Raman imaging demonstrates FGF2-induced craniosynostosis in mouse calvaria.

Study of localization of response to fibroblast growth factor-2 in murine calvaria using Raman spectroscopic imaging