Processing cell-seeded polyester scaffolds for histology

Holy, Chantal E.; Yakubovich, Raisa

doi:10.1002/(sici)1097-4636(200005)50:2<276::aid-jbm23>3.3.co;2-u

Cited by 1 publication

(2 citation statements)

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“…To confirm that bone had formed within the polymer scaffold, various histology stains were applied to fragments of a cell‐seeded PEMA disk maintained in culture for 5 weeks. Paraffin sections were not prepared because the organic solvents used for paraffin embedding would dissolve the polymer scaffold, resulting in a loss in morphological information 7. The red color observed for Alizarin red and Sirius red sections confirmed the deposition of mineral and collagen, respectively.…”

Section: Discussionmentioning

confidence: 99%

“…The resulting loss of three‐dimensional information is a serious limitation. Other problems include the dissolution of polymeric scaffolds by the organic solvents used in the preparation of thin sections7 or the disruption of early mineral deposits by aqueous solvents 8. A number of noninvasive optical tomography methods are available, but despite their high spatial resolution they have limited depth penetration and local area coverage, and are dependent on the use of fluorescent probes.…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%