title: Delayed fracture healing and increased callus adiposity in a C57BL/6J murine model of obesity-associated type 2 diabetes mellitus.
abstract: BACKGROUND
  Impaired healing and non-union of skeletal fractures is a major public health problem, with morbidity exacerbated in patients with diabetes mellitus (DM). DM is prevalent worldwide and affects approximately 25.8 million US adults, with >90% having obesity-related type 2 DM (T2DM). While fracture healing in type 1 DM (T1DM) has been studied using animal models, an investigation into delayed healing in an animal model of T2DM has not yet been performed.
  
  
  METHODS
  Male C57BL/6J mice at 5 weeks of age were placed on either a control lean diet or an experimental high-fat diet (HFD) for 12 weeks. A mid-diaphyseal open tibia fracture was induced at 17 weeks of age and a spinal needle was used for intra-medullary fixation. Mice were sacrificed at days 7, 10, 14, 21, 28, and 35 for micro-computed tomography (μCT), histology-based histomorphometry and molecular analyses, and biomechanical testing.
  
  
  RESULTS
  HFD-fed mice displayed increased body weight and impaired glucose tolerance, both characteristic of T2DM. Compared to control mice, HFD-fed mice with tibia fractures showed significantly (p<0.001) decreased woven bone at day 28 by histomorphometry and significantly (p<0.01) decreased callus bone volume at day 21 by μCT. Interestingly, fracture calluses contained markedly increased adiposity in HFD-fed mice at days 21, 28, and 35. HFD-fed mice also showed increased PPARγ immunohistochemical staining at day 14. Finally, calluses from HFD-fed mice at day 35 showed significantly (p<0.01) reduced torsional rigidity compared to controls.
  
  
  CONCLUSIONS
  Our murine model of T2DM demonstrated delayed fracture healing and weakened biomechanical properties, and was distinctly characterized by increased callus adiposity. This suggests altered mesenchymal stem cell fate determination with a shift to the adipocyte lineage at the expense of the osteoblast lineage. The up-regulation of PPARγ in fracture calluses of HFD-fed mice is likely involved in the proposed fate switching.
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; School of Medicine and Dentistry, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Brown
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Kiminori
authors.initials: K
authors.lastname: Yukata
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Farnsworth
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Biostatistics and Computational Biology, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Ding-Geng
authors.initials: DG
authors.lastname: Chen
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Biomechanical Engineering, University of Rochester, Rochester, New York, United States of America.
authors.forename: Hani
authors.initials: H
authors.lastname: Awad
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Hilton
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Regis J
authors.initials: RJ
authors.lastname: O'Keefe
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Lianping
authors.initials: L
authors.lastname: Xing
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Mooney
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, United States of America; Department of Orthopaedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Zuscik
mesh_terms: D050154:Adiposity; D000818:Animals; D001696:Biomechanical Phenomena; D002146:Bony Callus; D003924:Diabetes Mellitus, Type 2 / Q000209:etiology* / Q000473:pathology*; D059305:Diet, High-Fat; D004195:Disease Models, Animal; D017102:Fracture Healing; D050723:Fractures, Bone; D015870:Gene Expression; D018149:Glucose Intolerance; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic; D009765:Obesity / Q000150:complications* / Q000209:etiology; D009929:Organ Size; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D055114:X-Ray Microtomography
chemical_list: D047495:PPAR gamma