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Ex-vivo Model of Breast Cancer Bone Metastasis. Keagan Collins, Laura Graefe, Chris Hubley, Jaymin Modi, Bethany Porter, & Ian Roberts. Sponsored by Nohe Lab, Department of Biological Sciences, University of Delaware. Background & Significance.

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ex vivo model of breast cancer bone metastasis

Ex-vivo Model of Breast Cancer Bone Metastasis

Keagan Collins, Laura Graefe, Chris Hubley, JayminModi, Bethany Porter, & Ian Roberts

Sponsored by Nohe Lab, Department of Biological Sciences, University of Delaware

background significance
Background & Significance
  • Dr. Anja Nohe is an Assistant Professor in the Department of Biological Sciences at UD
  • Interested in metastasizing breast cancer and maintaining cell viability within a femoral head
  • Current treatments and diagnostic capabilities for metastasized breast cancer are not very effective and research into the field stands to change the disease
project scope
Project Scope
  • To develop a perfusion bioreactor that maintains cell viability within a femoral head with the purpose of metastasizing breast cancer cells
wants constraints
Wants & Constraints

Constraints

Wants

  • Bone remains viable for at least ten days
  • Project deadline
  • Sterility of bioreactor
  • Ability to test for cell viability
  • Integrity of vasculature
  • Ease of use
  • Efficient use of resources
  • Ideal size
  • Cost effective
cannulation
Cannulation
  • Two possible methods
    • Through existing vasculature
    • Through drilled entry points
  • Dependent on individual bone sample
bone type
Bone Type
  • Originally adult bovine femoral head
    • Bone cells were no longer viable
  • First trial used juvenile bovine (veal) femoral heads
    • Bone will be acquired within 24 hours
    • Many cells should remain viable
final design concept1
Final Design Concept

Bioreactor Chamber

Vibration Table

testing
Testing
  • Cell Viability
    • Glucose testing
    • pH testing
  • Contamination
  • Cannulation/perfusion
  • Vibration table

Glucose and pH testing

trial two results
Trial Two Results

Approximately 99% initial cell viability

92% viability maintained in 12 hours

failure analysis
Failure Analysis
  • Possible methods of failure
    • Contamination
    • Initial cell viability
    • Catheter placement
    • Media viability testing

Media samples taken every 36 hours

design evaluation
Design Evaluation
  • Major metrics met
    • Viability maintained
    • Vasculature intact
    • Mechanical loading conditions
    • No apparent bacterial contamination
  • Complications
    • Fungal contamination
path forward
Path Forward
  • Full length 10 day trial with human femoral head
  • Full length 10 day trial during which metastatic breast cancer cells are introduced into human femoral head
  • Study of effects of metastatic breast cancer on human femoral heads
references
References

[1] Carreau, A., B. El Hafny-Rahbi, A. Matejuk, C. Grillion, and C. Kieda. "Why Is the Partial Oxygen Pressure of Human Tissues a Crucial Parameter? Small Molecules and Hypoxia." J Cell Molecular Medicine (2011): 1239-253. Pubmed. Web.

[2] Kellum, John A. "Determinants of Blood PH in Health and Disease." Determinants of Blood PH in Health and Disease (2000): 6-14. Pubmed. Web. 3 Sept. 2013.

[3] De Oliveira, Mônica Longo, Cássia T. Bergamaschi, Orivaldo Lopes Silva, Keiko Okino Nonaka, Charles Chenwei Wang, Aluízio Barbosa Carvalho, Vanda Jorgetti, Ruy R. Campos, and Marise Lazaretti-Castro. "Mechanical Vibration Preserves Bone Structure in Rats Treated with Glucocorticoids." Bone 46.6 (2010): 1516-521. Print.

[4] Kubo, T., K. Kimori, F. Nakamura, and S. Inoue. "Blood Flow and Blood Volume in the Femoral Heads of Healthy Adults According to Age: Measurement with Positron Emission Tomography (PET)." Ann Nucleic Medicine (2001): 231-35. Pubmed. Web. 3 Sept. 2013.

[5] Fisher, J.P., & Yeatts, A.B. (2011).  Bone tissue engineering bioreactors: Dynamic culture and the influence of shear stress.  Bone, 48 (2), 171-181

[6]Metastatic Breast Cancer. (n.d.). Patient Resource. Retrieved September 6, 2013, from http://www.patientresource.com/Metastatic_Breast.aspx

special thanks to
Special thanks to…
  • Advisor: Dr. Liyun Wang, Department of Mechanical Engineering
  • Sponsor: Dr. AnjaNohe, Department of Biological Sciences
  • HemanthAkkiraju and Jeremy Bonorin the Nohe Lab
  • Dr. Gwen Talham, Department of Animal Care
  • Dr. Robert Sikes, Director of Center for Translational Cancer Research, Department of Biological Sciences