Category Research project
  • Alternativmethoden zum Tierversuch

Deferoxamine (DFO/Desferal®) to accelerate bone healing and treat fracture healing disorders

Project status
Completed
Project start
Mar 2021
Project end
Feb 2024
Acronym
PROOF-DESFERAL
Department
Experimentelle Toxikologie und ZEBET

Description and Objective

In the proposed study, we aim at paving the path for using the HIF-stabilizer Deferoxamine (DFO) for a new application to prevent fracture healing disorders as a cost-effective and low-risk alternative to recombinant growth factors such as BMP-2. We consider the FDAshort forFood and Drug Administration approved DFO and commercially available as Desferal® (Novartis), listed on World Health Organization's List of Essential Medicines, suitable for rapid clinical translation to improve fracture healing and the treatment of bone healing disorders. In the next step, we strive for clinical translation. The BfRshort forGerman Federal Institute for Risk Assessment is participating in this study as one of two laboratories that will test varying concentrations of DFO regarding their influence on mesenchymal stromal cells (precursor cells for bone forming osteoblasts). Both laboratories will work with harmonized protocols and the same cell culture reagents and cells. Furthermore, both laboratories will receive blinded samples of DFO in different concentrations. The data from both laboratories will then be send to a third project partner for evaluation. This is meant to investigate how good the comparability of in vitro studies from different laboratories actually is and if such study design has clear advantages. We also plan to investigate the effect of DFO in our bioreactor (bone-on-a-chip). In addition, the data from the in vitro experiment is meant to be compared with data from animal experiments to evaluate how precise a carfully conducted in vitro study can predict the outcome of in vivo approaches. Data from the bone-on-a-chip experiments will also be compared to results from in vitro cell culture and in vivo experiments to classify their significance. These research questions have direct relevance for the work of Department 9 (Experimental Toxicology and ZEBET) in the field of the 3R (Reduce, Replace, Refine), since this study is meant to show that the number of animal experiments can be reduced by an improved study design and the use of reliable in vitro

Result

In the present study, the goal was to introduce Desferoxamine (DFO; Desferal®, Novartis) as a stabilizer of hypoxia-inducible factor (HIF) to prevent fracture healing disorders, offering a cost-effective and low-risk alternative to other treatment options, such as the administration of growth factors, in order to improve fracture healing and prevent bone healing disorders in clinical practice. The objective of subproject 01KC2011B was to conduct two-dimensional (2D) and three-dimensional (3D) in vitro experiments to assess the extent to which DFO enhances osteogenic differentiation/calcification.For this purpose, 2D and 3D (spheroid) cultures of primary human mesenchymal stromal cells from 26 (2D) and 16 (3D) donors were established and treated for 21 days with different concentrations of DFO (2D = blinded; 3D 0/250 µM) in osteogenic differentiation medium (ODM). While the 2D culture showed a changed mineralization rate under the different treatments (concentrations of DFO), the specific assignment of these changes will only be determined in the final report after unblinding by a collaborating laboratory. The 3D culture, however, did not show any differences in mineralization under the different conditions (0 µM and 250 µM DFO, each in ODM) compared to the normal medium (NM) control. The additional determination of alkaline phosphatase activity in the 3D culture indicated a (non-significant) increase in enzyme activity under DFO treatment compared to the NM control. Additionally, an increase in DNA content was observed in these samples due to culture in ODM (0 µM and 250 µM DFO). The classification of these results with regard to follow-up experiments in mice and sheep was carried out in collaboration with the other project partners.
Type of project

Third-party funded project

Research focus

Gesundheit von Mensch, Tier und Umwelt (One Health) / Alternativmethoden zum Tierversuch

Organisational units and partners

Lead specialist group: Zentralstelle zur Erfassung und Bewertung von Ersatz- und Ergänzungsmethoden zum Tierversuch (91)
Contact persons: Dr. Elisa Wistorf, Dr. Julia Scheinpflug
External partner: Charité - Universitätsmedizin Berlin, Deutsches Rheuma-Forschungszentrum Berlin, Tierklinik Bad Langensalza, Forschungszentrum für Medizintechnik und Biotechnologie, Zentrum für Traumaforschung

Funding body and grant number

Bundesministerium für Bildung und Forschung
01KC2011B