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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, 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

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  1. 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

  2. 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

  3. Project Scope • To develop a perfusion bioreactor that maintains cell viability within a femoral head with the purpose of metastasizing breast cancer cells

  4. 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

  5. Biological Metrics

  6. Additional Metrics

  7. Cannulation • Two possible methods • Through existing vasculature • Through drilled entry points • Dependent on individual bone sample

  8. 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

  9. Final Design Concept

  10. Final Design Concept Bioreactor Chamber Vibration Table

  11. Cost Analysis

  12. Initial Trial - Bovine

  13. Initial Trial - Bovine

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

  15. Trial One Results

  16. Trial One Results

  17. Trial Two Results Approximately 99% initial cell viability 92% viability maintained in 12 hours

  18. Failure Analysis • Possible methods of failure • Contamination • Initial cell viability • Catheter placement • Media viability testing Media samples taken every 36 hours

  19. Design Evaluation • Major metrics met • Viability maintained • Vasculature intact • Mechanical loading conditions • No apparent bacterial contamination • Complications • Fungal contamination

  20. 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

  21. 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

  22. 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

  23. Questions?

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