title: Localized deferoxamine injection augments vascularity and improves bony union in pathologic fracture healing after radiotherapy.
abstract: BACKGROUND
  Medically based efforts and alternative treatment strategies to prevent or remediate the corrosive effects of radiotherapy on pathologic fracture healing have failed to produce clear and convincing evidence of success. Establishing an effective pharmacologic option to prevent or treat the development of non-unions in this setting could have immense therapeutic potential. Experimental studies have shown that deferoxamine (DFO), an iron-chelating agent, bolsters vascularity and subsequently enhances normal fracture healing when injected locally into a fracture callus in long bone animal models. Since radiotherapy is known to impede angiogenesis, we hypothesized that the pharmacologic addition of DFO would serve to mitigate the effects of radiotherapy on new vessel formation in vitro and in vivo.
  
  
  METHODS
  In vitro investigation of angiogenesis was conducted utilizing HUVEC cells in Matrigel. Endothelial tubule formation assays were divided into four groups: Control, Radiated, Radiated+Low-Dose DFO and Radiated+High-Dose DFO. Tubule formation was quantified microscopically and video recorded for the four groups simultaneously during the experiment. In vivo, three groups of Sprague-Dawley rats underwent external fixator placement and fracture osteotomy of the left mandible. Two groups received pre-operative fractionated radiotherapy, and one of these groups was treated with DFO after fracture repair. After 40 days, the animals were perfused and imaged with micro-CT to calculate vascular radiomorphometrics.
  
  
  RESULTS
  In vitro, endothelial tubule formation assays demonstrated that DFO mitigated the deleterious effects of radiation on angiogenesis. Further, high-dose DFO cultures appeared to organize within 2h of incubation and achieved a robust network that was visibly superior to all other experimental groups in an accelerated fashion. In vivo, animals subjected to a human equivalent dose of radiotherapy (HEDR) and left mandibular fracture demonstrated quantifiably diminished μCT metrics of vascular density, as well as a 75% incidence of associated non-unions. The addition of DFO in this setting markedly improved vascularity as demonstrated with 3D angiographic modeling. In addition, we observed an increased incidence of bony unions in the DFO treated group when compared to radiated fractures without treatment (67% vs. 25% respectively).
  
  
  CONCLUSIONS
  Our data suggest that selectively targeting angiogenesis with localized DFO injections is sufficient to remediate the associated severe vascular diminution resulting from a HEDR. Perhaps the most consequential and clinically relevant finding was the ability to reduce the incidence of non-unions in a model where fracture healing was not routinely observed.
authors.affiliation: Craniofacial Research Laboratory, Plastic Surgery Section, University of Michigan, Ann Arbor, MI 48109-4215, USA.
authors.forename: Alexis
authors.initials: A
authors.lastname: Donneys
authors.forename: Daniela M
authors.initials: DM
authors.lastname: Weiss
authors.forename: Sagar S
authors.initials: SS
authors.lastname: Deshpande
authors.forename: Salman
authors.initials: S
authors.lastname: Ahsan
authors.forename: Catherine N
authors.initials: CN
authors.lastname: Tchanque-Fossuo
authors.forename: Deniz
authors.initials: D
authors.lastname: Sarhaddi
authors.forename: Benjamin
authors.initials: B
authors.lastname: Levi
authors.forename: Steven A
authors.initials: SA
authors.lastname: Goldstein
authors.forename: Steven R
authors.initials: SR
authors.lastname: Buchman
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003676:Deferoxamine / Q000008:administration & dosage*; D017102:Fracture Healing; D006801:Humans; D009389:Neovascularization, Pathologic; D011878:Radiotherapy; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D003676:Deferoxamine