title: The role of the adaptive immune system in burn-induced heterotopic ossification and mesenchymal cell osteogenic differentiation.
abstract: Heterotopic ossification (HO) is the pathologic process of extraskeletal bone formation. Although the exact etiology remains unknown, inflammation appears to catalyze disease progression. The goal of this study is to determine the impact of the adaptive immune system on HO.
  
  
  
  HO was induced in 8-wk-old control C57BL/6 and immunocompromised Rag1tm1Mom (Rag1 KO) male mice deficient in B- and T-lymphocytes via combined Achilles tenotomy and burn injury. Microcomputed tomography quantified the extent of HO formation at the tenotomy site. Adipose-derived mesenchymal stem cells were harvested to evaluate osteogenic differentiation potential.
  
  
  
  Areas of developing HO demonstrated substantial enrichment of CD45 + leukocytes at 3 wk after injury. HO from Rag1 KO mice was substantially less mature with foci of cartilage and disorganized trabecular bone present 12 wk after injury. Rag1 KO mice formed 60% less bone compared to immunocompetent controls (4.67 ± 1.5 mm versus 7.76 ± 0.65 mm; P = 0.001). Tartrate-resistant acid phosphatase staining and immunofluorescent analysis of osteoprotegerin and nuclear factor kappa-light-chain-enhancer of activated B cells demonstrated no appreciable difference in osteoclast number or activation. Alizarin red staining in vitro demonstrated a significant decrease in osteogenic potential in immunocompromised mice compared to controls (29.1 ± 0.54 mm versus 12.1 ± 0.14 mm; P < 0.001).
  
  
  
  We demonstrate a prominent role for the adaptive immune system in the development of HO. In the absence of mature B- and T-lymphocytes, HO growth and development are attenuated. Furthermore, we demonstrate that mesenchymal populations from B- and T-cell deficient mice are inherently less osteogenic. This study identifies a potential therapeutic role for modulation of the adaptive immune system in the treatment of HO.
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Kavitha
authors.initials: K
authors.lastname: Ranganathan
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Shailesh
authors.initials: S
authors.lastname: Agarwal
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: David
authors.initials: D
authors.lastname: Cholok
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Shawn
authors.initials: S
authors.lastname: Loder
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Hsiao Hsin
authors.initials: HH
authors.lastname: Sung Hsieh
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Stewart C
authors.initials: SC
authors.lastname: Wang
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Buchman
authors.affiliation: Department of Surgery, University of Michigan Health Systems, Ann Arbor, Michigan. Electronic address: blevi@med.umich.edu.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Levi
mesh_terms: D056704:Adaptive Immunity; D000818:Animals; D002056:Burns / Q000150:complications* / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000276:immunology / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009999:Ossification, Heterotopic / Q000000981:diagnostic imaging / Q000209:etiology* / Q000276:immunology; D010012:Osteogenesis / Q000276:immunology*; D055114:X-Ray Microtomography
keywords: Adaptive immune system; Bone biology; Ectopic bone; Heterotopic ossification; Trauma