title: Mechanistic insight into the effects of Aryl Hydrocarbon Receptor activation on osteogenic differentiation.
abstract: While inhibition of bone healing and increased rates of pseudarthrosis are known adverse outcomes associated with cigarette smoking, the underlying mechanisms by which this occurs are not well understood. Recent work has implicated the Aryl Hydrocarbon Receptor (Ahr) as one mediator of the anti-osteogenic effects of cigarette smoke (CS), which contains numerous toxic ligands for the Ahr. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) is a high-affinity Ahr ligand frequently used to evaluate Ahr pathway activation. The purpose of this study was to elucidate the downstream mechanisms of dioxin action on bone regeneration and investigate Ahr antagonism as a potential therapeutic approach to mitigate the effects of dioxin on bone. Markers of osteogenic activity and differentiation were assessed in primary rat bone marrow stromal cells (BMSC) after exposure to dioxin, Ahr antagonists, or antagonist + dioxin. Four Ahr antagonists were evaluated: α-Naphthoflavone (ANF), resveratrol (Res), 3,3'-Diindolylmethane (DIM), and luteolin (Lut). Our results demonstrate that dioxin inhibited ALP activity, migratory capacity, and matrix mineralization, whereas co-treatment with each of the antagonists mitigated these effects. Dioxin also inhibited BMSC chemotaxis, while co-treatment with several antagonists partially rescued this effect. RNA and protein expression studies found that dioxin down-regulated numerous pro-osteogenic targets, whereas co-treatment with Ahr antagonists prevented these dioxin-induced expression changes to varying degrees. Our results suggest that dioxin adversely affects bone regeneration in a myriad of ways, many of which appear to be mediated by the Ahr. Our work suggests that the Ahr should be investigated as a therapeutic target to combat the adverse effects of CS on bone healing.
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Chawon
authors.initials: C
authors.lastname: Yun
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Weiner
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Danielle S
authors.initials: DS
authors.lastname: Chun
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Jonghwa
authors.initials: J
authors.lastname: Yun
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Ralph W
authors.initials: RW
authors.lastname: Cook
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Schallmo
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Abhishek S
authors.initials: AS
authors.lastname: Kannan
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Sean M
authors.initials: SM
authors.lastname: Mitchell
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Ryan D
authors.initials: RD
authors.lastname: Freshman
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Park
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Wellington K
authors.initials: WK
authors.lastname: Hsu
authors.affiliation: Northwestern University Department of Orthopaedic Surgery, Chicago, IL, USA.
authors.forename: Erin L
authors.initials: EL
authors.lastname: Hsu
keywords: Aryl Hydrocarbon Receptor; Bone regeneration; Cigarette smoke; Dioxin; TCDD