gms | German Medical Science

Deutscher Kongress für Orthopädie und Unfallchirurgie (DKOU 2016)

25.10. - 28.10.2016, Berlin

An orthotopic humanized bone model for preclinical bone tumor research

Meeting Abstract

  • presenting/speaker Ferdinand Wagner - Institute of Health and Biomedical Innovation, Queensland University of Technology, Orthopädische Klinik der Universität Regensburg, Brisbane, Australia
  • Boris M. Holzapfel - Queensland University of Technology, Orthopädische Klinik der Universität Würzburg, Brisbane, Australia
  • Laure Martine - Institute of Health and Biomedical Innovations, Queensland University of Technology, Kelvin Grove, Australia
  • Jeremy Baldwin - Institute of Health and Biomedical Innovations, Queensland University of Technology, Kelvin Grove, Australia
  • Felix Wunner - Institute of Health and Biomedical Innovations, Queensland University of Technology, Kelvin Grove, Australia
  • Cosmo Orlando Hutmacher - Institute of Health and Biomedical Innovations, Queensland University of Technology, Kelvin Grove, Australia
  • Daniela Lössner - Institute of Health and Biomedical Innovations, Queensland University of Technology, Kelvin Grove, Australia
  • Dietmar Werner Hutmacher - Institute of Health and Biomedical Innovations, Queensland University of Technology, Kelvin Grove, Australia

Deutscher Kongress für Orthopädie und Unfallchirurgie (DKOU 2016). Berlin, 25.-28.10.2016. Düsseldorf: German Medical Science GMS Publishing House; 2016. DocGR13-868

doi: 10.3205/16dkou416, urn:nbn:de:0183-16dkou4165

Veröffentlicht: 10. Oktober 2016

© 2016 Wagner et al.
Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung). Lizenz-Angaben siehe http://creativecommons.org/licenses/by/4.0/.


Gliederung

Text

Objectives: Humanized xenograft models try to minimize interspecies-specific differences between murine host organisms and the investigated human tumor by engineering a human tissue microenvironment. This enables more predictable translation of experimental results from bench to bedside. We questioned if an orthotopic humanized tissue engineered bone construct (ohTEBC) at the mouse femur can be utilized as a suitable niche for osteosarcoma (OS) growth.

Methods: A biodegradable polycaprolactone (PCL) tubular scaffold was generated by electro-melt spinning and seeded with human osteogenically differentiated mesenchymal progenitor cells. The scaffolds where then implanted at the femur of NOD/SCID mice together with recombinant human bone morphogenetic protein 7 (rhBMP-7) embedded in a fibrin glue matrix. Six weeks after implantation luciferase labeled human osteosarcoma cells (Luc-SAOS-2) where injected directly into the constructs. Tumor growth was followed for 9 weeks via bioluminescence imaging. Tissues were harvested for bone morphology and tumor analysis.

Results and Conclusion: 6 weeks after implantion of ohTEBCs orthotopic bone formation was seen. After harvesting the femurs micro computed tomography showed that ohTEBCs have developed into an organ, which had all radiological features of bone. Around the original mouse femur new bone trabeculae have formed which were surrounded by a bone cortex. Hematoxilin and eosin (H&E) staining showed that the new trabecular network spaces were filled with bone marrow. By immunohistochemical staining for CD34 these cells could be verified as hematopoitic stem cells (HSCs). Staining for human specific (hs) collagen type-I (hs Col-I) showed that there was human extracellular bone matrix production. The presence of nuclei staining positive for human nuclear mitotic apparatus protein 1 (hs NuMa) proved the osteocytes residing within the bone matrix were also of human origin.

9 weeks after injection of Luc-SAOS-2 cells not only a primary tumor but also lung metastasis could be found via BLI. After euthanization histological analysis showed that these tumors produced bone, extracellular matrix and especially osteoid and therefore represented an osteoblastic subtype of OS. Positive hs NuMa staining depicted human tumor cells that produced human extracellular matrix as shown by hs Col-I. Ki67 staining showed the proliferative nature of the tumor tissue.

In Conclusion, this model can be utilized as a platform for primary bone tumor research. Within this humanized microenvironment OS growth developed within a small number of weeks and also produced clinical relevant lung metastases. The tumors showed large similarities to the human disease and might be favorable to the conventional in vivo model when investigating intercellular interactions of tumor cells with their surrounding microenvironment. OhTEBC also might be utilized for research of other bone malignancies like bone metastases from prostate and breast cancer as well a leukemia and multiple myeloma.