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Flow perfusion maintains ex vivo bone viability: a novel model for bone biology research.

Publication ,  Journal Article
Davidson, EH; Reformat, DD; Allori, A; Canizares, O; Janelle Wagner, I; Saadeh, PB; Warren, SM
Published in: J Tissue Eng Regen Med
November 2012

Encased in lacunae, osteocytes receive nutrition and biomechanical signals through the lacunocanalicular system. We have developed a novel flow-perfusion bioreactor designed to support lacunocanalicular fluid flow. We hypothesize that ex vivo fluid flow can maintain endochondral bone viability and, ultimately, serve as a novel model to study bone biology in vitro. Sprague-Dawley rat femurs were harvested, stripped of soft tissue, loaded into a custom-designed bioreactor and perfused with osteogenic culture medium. After 14 days of flow-perfusion or static culture, the bones were harvested, fixed, decalcified, embedded, sectioned and stained with haematoxylin and eosin. Fresh long bone samples were similarly processed for comparison. Osteocyte viability and function were also evaluated, using thiazolyl blue tetrazolium bromide (MTT), fluorospectrophotometric DNA quantification, alkaline phosphatase (ALP) colorimetric assay and fluorochrome labelling of mineralizing surfaces. All samples remained free of infection throughout the study period. After 14 days of flow perfusion, histological analysis showed normal-appearing bony architecture, with 72% of lacunae being osteocyte-filled compared with 93% in freshly harvested samples and only 36% in static samples. MTT staining and assay confirmed osteocyte viability in the flow-perfusion samples as well as in fresh samples. DNA quantification demonstrated DNA to be preserved in flow-perfused samples when compared with freshly harvested samples. ALP activity in flow-perfusion explants was upregulated compared with fresh and static samples. Fluorochrome-labelled mineralizing surfaces were seen throughout the explanted flow-perfused samples. This is the first demonstration that flow perfusion provides adequate chemotransportation to explanted murine endochondal bones.

Duke Scholars

Published In

J Tissue Eng Regen Med

DOI

EISSN

1932-7005

Publication Date

November 2012

Volume

6

Issue

10

Start / End Page

769 / 776

Location

England

Related Subject Headings

  • Transplants
  • Time Factors
  • Rats, Sprague-Dawley
  • Rats
  • Perfusion
  • Osteocytes
  • Organ Culture Techniques
  • Femur
  • DNA
  • Cell Survival
 

Citation

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Davidson, E. H., Reformat, D. D., Allori, A., Canizares, O., Janelle Wagner, I., Saadeh, P. B., & Warren, S. M. (2012). Flow perfusion maintains ex vivo bone viability: a novel model for bone biology research. J Tissue Eng Regen Med, 6(10), 769–776. https://doi.org/10.1002/term.478
Davidson, Edward H., Derek D. Reformat, Alessandro Allori, Orlando Canizares, I. Janelle Wagner, Pierre B. Saadeh, and Stephen M. Warren. “Flow perfusion maintains ex vivo bone viability: a novel model for bone biology research.J Tissue Eng Regen Med 6, no. 10 (November 2012): 769–76. https://doi.org/10.1002/term.478.
Davidson EH, Reformat DD, Allori A, Canizares O, Janelle Wagner I, Saadeh PB, et al. Flow perfusion maintains ex vivo bone viability: a novel model for bone biology research. J Tissue Eng Regen Med. 2012 Nov;6(10):769–76.
Davidson, Edward H., et al. “Flow perfusion maintains ex vivo bone viability: a novel model for bone biology research.J Tissue Eng Regen Med, vol. 6, no. 10, Nov. 2012, pp. 769–76. Pubmed, doi:10.1002/term.478.
Davidson EH, Reformat DD, Allori A, Canizares O, Janelle Wagner I, Saadeh PB, Warren SM. Flow perfusion maintains ex vivo bone viability: a novel model for bone biology research. J Tissue Eng Regen Med. 2012 Nov;6(10):769–776.
Journal cover image

Published In

J Tissue Eng Regen Med

DOI

EISSN

1932-7005

Publication Date

November 2012

Volume

6

Issue

10

Start / End Page

769 / 776

Location

England

Related Subject Headings

  • Transplants
  • Time Factors
  • Rats, Sprague-Dawley
  • Rats
  • Perfusion
  • Osteocytes
  • Organ Culture Techniques
  • Femur
  • DNA
  • Cell Survival