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 • 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 • To develop a perfusion bioreactor that maintains cell viability within a femoral head with the purpose of metastasizing breast cancer cells Wants & Constraints Constraints Wants 1. Bone remains viable for at least ten days 2. Project deadline 3. Sterility of bioreactor 4. Ability to test for cell viability 5. Integrity of vasculature 1. Ease of use 2. Efficient use of resources 3. Ideal size 4. Cost effective Biological Metrics Additional Metrics Cannulation • Two possible methods ▫ Through existing vasculature ▫ Through drilled entry points • Dependent on individual bone sample 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 Concept Final Design Concept Bioreactor Chamber Vibration Table Cost Analysis Initial Trial - Bovine Initial Trial - Bovine Testing • Cell Viability ▫ Glucose testing ▫ pH testing • Contamination • Cannulation/perfusion • Vibration table Glucose and pH testing Trial One Results Glucose Usage (mg/h) at a function of Time 140 Glucose Usage Rate (mg/h) 120 100 80 60 40 20 0 0 50 100 150 Time (hours) 200 250 Trial One Results Bioreactor Media pH as a Function of Time 9 8.5 8 pH 7.5 7 6.5 6 5.5 5 0 50 100 150 Time (hours) 200 250 Trial Two Results Approximately 99% initial cell viability 92% viability maintained in 12 hours Failure Analysis • Possible methods of failure ▫ ▫ ▫ ▫ Contamination Initial cell viability Catheter placement Media viability testing Media samples taken every 36 hours Design Evaluation • Major metrics met ▫ Viability maintained ▫ Vasculature intact ▫ Mechanical loading conditions ▫ No apparent bacterial contamination • Complications ▫ Fungal contamination 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  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.  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.  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.  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.  Fisher, J.P., & Yeatts, A.B. (2011). Bone tissue engineering bioreactors: Dynamic culture and the influence of shear stress. Bone, 48 (2), 171-181 Metastatic Breast Cancer. (n.d.). Patient Resource. Retrieved September 6, 2013, from http://www.patientresource.com/Metastatic_Breast.aspx Special thanks to… • Advisor: Dr. Liyun Wang, Department of Mechanical Engineering • Sponsor: Dr. Anja Nohe, Department of Biological Sciences • Hemanth Akkiraju and Jeremy Bonor in the Nohe Lab • Dr. Gwen Talham, Department of Animal Care • Dr. Robert Sikes, Director of Center for Translational Cancer Research, Department of Biological Sciences Questions?