Bone is a complex system consisting of a carbonated apatitic calcium phosphate phase supported on a collagen-rich extracellular matrix. While the macroscopic properties of bone have been extensively investigated using different techniques in various biological systems, the early steps in bone tissue mineralization are not well understood. In an effort to characterize bone growth, numerous studies have been conducted to understand the chemical mechanisms involved in both de novo bone formation and bone remodeling. Despite this increased interest, an understanding of the processes governing early mineralization of bone is still incomplete. This proposal will use near-infrared Raman microspectroscopy and Raman imaging to study early mineralization in the mouse calvaria (the flat bones that comprise the top of the skull). The goal is to understand the steps leading to the deposition of new mineral and the chemical and physical transformations that the mineral undergoes during fetal and early postnatal development. Because Raman microspectroscopy is a relatively new tool in bone research, the work will begin with validation of protocols for Raman spectroscopy of bone tissue. The effects of standard specimen fLxing and embedding protocols and the potential Raman or fluorescence interference from a set of common histological stains will be examined. It is proposed that there will be few or no problems encountered beyond the known effects, such as protein cross-linking, of certain fixing protocols. If interferences due to the staining protocols occur, multivariate spectral and image processing will be used to overcome them. Because the progress of mineralization is accompanied by shifts in mineral spectral bands, the interpretation of small spectral band shifts will be put on a fawn foundation. In a subcontract to Central Michigan University, vibrational spectra (band positions and frequencies) of substituted hydroxyapatites will be computed by density functional theory and compared to Raman spectral measurements of model compounds (synthetic non-substituted and substituted hydroxyapatites) and on murine calvarial bone tissue. Systematic Raman imaging will be performed on calvarial sections harvested from normal mice just prior to the onset of mineralization (fetal day 15.5) until postnatal day 14. Raman imaging will be carried out at 1-day intervals during the prenatal period and at 2-day intervals during the postnatal period. Correlative immunohistological staining and Raman imaging will be used to understand the effects of bone sialoprotein and osteocalcin on the course of mineralization.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR047969-04
Application #
6986838
Study Section
Special Emphasis Panel (ZRG1-BECM (01))
Program Officer
Lester, Gayle E
Project Start
2003-01-10
Project End
2007-11-30
Budget Start
2005-12-01
Budget End
2007-11-30
Support Year
4
Fiscal Year
2006
Total Cost
$295,489
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Zhang, Yanshuai; McNerny, Erin Gatenby; Terajima, Masahiko et al. (2016) Loss of BMP signaling through BMPR1A in osteoblasts leads to greater collagen cross-link maturation and material-level mechanical properties in mouse femoral trabecular compartments. Bone 88:74-84
Addison, W N; Nelea, V; Chicatun, F et al. (2015) Extracellular matrix mineralization in murine MC3T3-E1 osteoblast cultures: an ultrastructural, compositional and comparative analysis with mouse bone. Bone 71:244-56
Tchanque-Fossuo, Catherine N; Gong, Bo; Poushanchi, Behdod et al. (2013) Raman spectroscopy demonstrates Amifostine induced preservation of bone mineralization patterns in the irradiated murine mandible. Bone 52:712-717
Esmonde-White, Karen A; Esmonde-White, Francis W L; Holmes, Crystal M et al. (2013) Alterations to bone mineral composition as an early indication of osteomyelitis in the diabetic foot. Diabetes Care 36:3652-4
McElderry, John-David P; Zhao, Guisheng; Khmaladze, Alexander et al. (2013) Tracking circadian rhythms of bone mineral deposition in murine calvarial organ cultures. J Bone Miner Res 28:1846-54
Meganck, J A; Begun, D L; McElderry, J D et al. (2013) Fracture healing with alendronate treatment in the Brtl/+ mouse model of osteogenesis imperfecta. Bone 56:204-12
McElderry, John-David P; Zhu, Peizhi; Mroue, Kamal H et al. (2013) Crystallinity and compositional changes in carbonated apatites: Evidence from 31P solid-state NMR, Raman, and AFM analysis. J Solid State Chem 206:
Demers, Jennifer-Lynn H; Davis, Scott C; Pogue, Brian W et al. (2012) Multichannel diffuse optical Raman tomography for bone characterization in vivo: a phantom study. Biomed Opt Express 3:2299-305
Esmonde-White, Francis W L; Esmonde-White, Karen A; Morris, Michael D (2011) Minor distortions with major consequences: correcting distortions in imaging spectrographs. Appl Spectrosc 65:85-98
McElderry, John-David P; Kole, Matthew R; Morris, Michael D (2011) Repeated freeze-thawing of bone tissue affects Raman bone quality measurements. J Biomed Opt 16:071407

Showing the most recent 10 out of 24 publications