Bioreactor and probe system for magnetic resonance microimaging and spectroscopy of chondrocytes and neocartilage

Petersen, Erik F.; Potter, Kimberlee; Butler, John J.; Fishbein, Kenneth W.; Horton, Walter E.; Spencer, Richard G.; McFarland, Eric W.

doi:10.1002/(sici)1098-1098(1997)8:3<285::aid-ima6>3.0.co;2-6

Cited by 22 publications

(37 citation statements)

References 3 publications

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“…The data presented here are an extension of results obtained previously with the same HFBR system. (12)(13)(14) The principal limitation of this system is that calcified cartilage does not remodel through subsequent stages leading to woven or lamellar bone.…”

Section: Discussionmentioning

confidence: 99%

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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Section: Discussionmentioning

confidence: 99%

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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“…Hollow-fiber bioreactors (HFBRs) provide a suitable model system for both of these goals, permitting the growth of macroscopic amounts of 3-dimensional hyaline cartilage (3,4) subject to a wide variety of tissuemodifying agents and interventions (5). In previous work, we have found that the neocartilage develops from isolated chondrocytes in the HFBR in a highly reproducible manner, permitting the study of cartilage formation, maintenance, and turnover (3)(4)(5).…”

mentioning

confidence: 99%

“…In previous work, we have found that the neocartilage develops from isolated chondrocytes in the HFBR in a highly reproducible manner, permitting the study of cartilage formation, maintenance, and turnover (3)(4)(5).…”

mentioning

confidence: 99%

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Matrix fixed‐charge density as determined by magnetic resonance microscopy of bioreactor‐derived hyaline cartilage correlates with biochemical and biomechanical properties

Chen¹,

Fishbein²,

Torzilli³

et al. 2003

Arthritis & Rheumatism

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Objective. To use noninvasive magnetic resonance imaging (MRI), biochemical analyses, and mechanical testing of engineered neocartilage grown in a hollowfiber bioreactor (HFBR) to establish tissue properties, and to test the hypothesis that MRI can be used to monitor biochemical and biomechanical properties of neocartilage.Methods. Chondrocytes from day 16 embryonic chick sterna were inoculated into an HFBR and maintained for up to 4 weeks with and without exposure to chondroitinase ABC. The fixed-charge density (FCD) of the cartilage was determined using the MRI gadolinium exclusion method. The sulfated glycosaminoglycan (S-GAG), hydroxyproline, and DNA contents were determined using biochemical procedures, while dynamic and equilibrium moduli were determined from mechanical indentation tests.Results. S-GAG content, tissue cross-sectional area, and equilibrium modulus of the neocartilage increased with development time. There was a gradient of S-GAG content across the length of control neocartilage at the 4-week time point, with higher values being found toward the inflow region. Exposure to chondroitinase ABC resulted in a decrease in tissue area, negative FCD, proteoglycan content, and equilibrium and dynamic moduli. The treated bioreactors displayed a lengthwise variation in S-GAG content, with higher values toward the outflow end. Linear correlations were established among FCD, proteoglycan content, and biomechanical properties.Conclusion. HFBR-derived neocartilage showed regional variation in S-GAG content under control conditions, and in the decrease of S-GAG in response to enzyme treatment. In addition, the results support the hypothesis that tissue parameters derived from MRI can be used to noninvasively monitor focal neocartilage formation and biochemical and biomechanical properties.

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“…Our mineralizing model is based on an HFBR design, previously described for chondrocytes (42), in which culture medium flows through the lumen of hollow fibers and osteoblasts attach to their outer surfaces. The bioreactors were constructed from high-purity glass tubing (o.d.…”

Section: Mineralizing Bioreactormentioning

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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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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Bioreactor and probe system for magnetic resonance microimaging and spectroscopy of chondrocytes and neocartilage

Cited by 22 publications

References 3 publications

Cartilage Calcification Studied by Proton Nuclear Magnetic Resonance Microscopy

Cartilage Calcification Studied by Proton Nuclear Magnetic Resonance Microscopy

Matrix fixed‐charge density as determined by magnetic resonance microscopy of bioreactor‐derived hyaline cartilage correlates with biochemical and biomechanical properties

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

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