Histomorphometry of the Embryonic Avian Growth Plate by Proton Nuclear Magnetic Resonance Microscopy

Potter, Kimberlee; Landis, William J.; Spencer, Richard G.

doi:10.1359/jbmr.2001.16.6.1092

Cited by 11 publications

(12 citation statements)

References 33 publications

(38 reference statements)

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“…Similar observations have been made for collagen in calcified cartilage (32), in embryonic bone (31), and in a tissue-engineered phalanx model (12). We attribute this result to an increase in the collagen content of the mineralized zone, possibly brought about by the dehydration of constituent collagen fibrils.…”

Section: Discussionsupporting

confidence: 86%

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Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR microspectroscopy

Chesnick

Avallone

Leapman

et al. 2007

Bone

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We present a three-dimensional mineralizing model based on a hollow fiber bioreactor (HFBR) inoculated with primary osteoblasts isolated from embryonic chick calvaria. Using non-invasive magnetic resonance microscopy (MRM), the growth and development of the mineralized tissue around the individual fibers were monitored over a period of nine weeks. Spatial maps of the water proton MRM properties of the intact tissue, with 78 μm resolution, were used to determine changes in tissue composition with development. Unique changes in the mineral and collagen content of the tissue were detected with high specificity by proton density (PD) and magnetization transfer ratio (MTR) maps, respectively. At the end of the growth period, the presence of a bone-like tissue was verified by histology and the formation of poorly crystalline apatite was verified by selected area electron diffraction and electron probe X-ray microanalysis. FTIR microspectroscopy confirmed the heterogeneous nature of the bone-like tissue formed. FTIR-derived phosphate maps confirmed that those locations with the lowest PD values contained the most mineral, and FTIR-derived collagen maps confirmed that bright pixels on MTR maps corresponded to regions of high collagen content. In conclusion, the spatial mapping of tissue constituents by FTIR microspectroscopy corroborated the findings of non-invasive MRM measurements and supported the role of MRM in monitoring the bone formation process in vitro.

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Section: Discussionsupporting

confidence: 86%

“…In fact, calibration curves have been derived for this MRM parameter and the collagen content of articular cartilage (27), engineered cartilage (28), and collagen gels (27,29,30). In this mineralizing system, collagen, the predominant organic constituent of bone, was assessed using MT ratio images (12,13,31,32).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR microspectroscopy

Chesnick

Avallone

Leapman

et al. 2007

Bone

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“…These data also are consistent with high k m values measured for periosteal bone, as compared with the cartilage matrix, in the avian growth plate. (3) The k m parameter is especially valuable because it provides information about the state of the collagen fibers throughout the mineralization process. Importantly, k m results support the notion that collagen enrichment is a prerequisite for mineralization.…”

Section: Potter Et Almentioning

confidence: 99%

“…(2) The application of proton nuclear magnetic resonance (NMR) microscopy now makes it pos-sible to circumvent some of these difficulties and to examine both the organic and the inorganic phases of calcified tissues by noninvasive means and without fixation, destructive sectioning, or histological staining. (3) This study was motivated by the need for a rapid bioanalytical technique that can assess the mineralization of long bones and the process of endochondral ossification. At present, the approaches to many analyses of the time course of bone formation and remodeling demand that bones be harvested at different time points and subjected to invasive techniques such as dissection, fixation, sectioning, staining, or other procedures required for histomorphometry, electron diffraction, and scanning and transmission electron microscopy (TEM).…”

Section: Introductionmentioning

confidence: 99%

“…(3,4) Although proton NMR images of bone yield very little signal, investigators have successfully assessed trabecular architecture and hence strength and fracture risk of cancellous bone using the high-intensity signal from the intervening bone marrow. (5)(6)(7)(8) A few groups also have applied solid-state imaging techniques to detect phosphorus-31 nuclei immobilized in the inorganic matrix of bone.…”

Section: Introductionmentioning

confidence: 99%

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Cartilage Calcification Studied by Proton Nuclear Magnetic Resonance Microscopy

Potter

Leapman

Basser

et al. 2002

Journal of Bone and Mineral Research

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A three-dimensional (3D) mineralizing culture system using hollow fiber bioreactors has been developed to study the early stages of endochondral ossification by proton nuclear magnetic resonance (NMR) microscopy. Chondrocytes harvested from the cephalic half of the sterna from 17-day-old chick embryos were terminally differentiated with 33 nM of retinoic acid for 1 week and mineralization was initiated by the addition of 1% ␤-glycerophosphate to the culture medium. Histological sections taken after 6 weeks of development in culture confirmed calcification of the cartilage matrix formed in bioreactors. Calcium to phosphorus ratios (

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Bone formation in polymeric scaffolds evaluated by proton magnetic resonance microscopy and X‐ray microtomography

Washburn

Weir

Anderson

et al. 2004

J Biomedical Materials Res

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Magnetic resonance microscopy (MRM) and X-ray microtomography (XMT) were used to investigate de novo bone formation in porous poly(ethyl methacrylate) (PEMA) scaffolds, prepared by a novel co-extrusion process. PEMA scaffolds were seeded with primary chick calvarial osteoblasts and cultured under static conditions for up to 8 weeks. Bone formation within porous PEMA scaffolds was confirmed by the application of histologic stains to intact PEMA disks. Disks were treated with Alizarin red to visualize calcium deposits and with Sirius red to visualize regions of collagen deposition. DNA analysis confirmed that cells reached confluence on the scaffolds after 7 weeks in static culture. The formation of bone in PEMA scaffolds was investigated with water proton MRM. Quantitative MRM maps of the magnetization transfer ratio (MTR) yielded maps of protein deposition, and magnetic resonance (MR) relaxation times (T1 and T2) yielded maps of mineral deposition. The location of newly formed bone and local mineral concentrations were confirmed by XMT. By comparing MRM and XMT data from selected regions-of-interest in one sample, the inverse relationship between the MR relaxation times and mineral concentration was validated, and calibration curves for estimating the mineral content of cell-seeded PEMA scaffolds from quantitative MRM images were developed.

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Histomorphometry of the Embryonic Avian Growth Plate by Proton Nuclear Magnetic Resonance Microscopy

Cited by 11 publications

References 33 publications

Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR microspectroscopy

Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR microspectroscopy

Cartilage Calcification Studied by Proton Nuclear Magnetic Resonance Microscopy

Bone formation in polymeric scaffolds evaluated by proton magnetic resonance microscopy and X‐ray microtomography

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