Smoking delays chondrogenesis in a mouse model of closed tibial fracture healing.
Smoking delays the healing process and increases morbidity associated with many common musculoskeletal disorders, including long bone fracture. In the current study, a murine model of tibial fracture healing was used to test the hypothesis that smoking delays chondrogenesis after fracture. Mice were divided into two groups, a nonsmoking control group and a group exposed to cigarette smoke for 1 month prior to surgical tibial fracture. Mice were euthanized at 7, 14, and 28 days after surgery. The outcomes measured were immunohistochemical staining for type II collagen protein expression as a marker of cartilage matrix and proliferating cell nuclear antigen (PCNA) staining to measure proliferation at the site of injury. Toluidine blue staining and histomorphometry were used to quantify areas of cartilaginous and noncartilaginous fracture callus. Radiographs were analyzed for evidence of remodeling after injury. At day 7 after injury, mice exposed to cigarette smoke had a smaller fracture callus with less cartilage matrix compared to controls. Proliferation was present at high levels in both groups at this time point, but proliferating cells had a more immature morphology in the smoking group. At day 14, chondrogenesis was more active in smokers compared to controls, while a higher percentage of bone was present in the control animals. At day 28, X-ray analysis revealed a larger fracture callus remaining in the smoking animals. Together, these findings show that the chondrogenic phase of tibial fracture healing is delayed by smoking. This study represents, to our knowledge, the first analysis of molecular and cellular mechanisms of healing in a smoking mouse fracture model.
El-Zawawy
Hossam B
HB
Department of Orthopaedic Surgery, Washington University School of Medicine at Barnes-Jewish Hospital, 660 S. Euclid Avenue, Campus Box 8233, St. Louis, Missouri 63110, USA.
Gill
Corey S
CS
Wright
Rick W
RW
Sandell
Linda J
LJ
D000818:Animals; D020219:Chondrogenesis / Q000187:drug effects*; D024043:Collagen Type II / Q000096:biosynthesis; D004195:Disease Models, Animal; D017102:Fracture Healing / Q000187:drug effects*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011859:Radiography; D012907:Smoking / Q000009:adverse effects*; D013194:Staining and Labeling; D013978:Tibial Fractures / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*; D014048:Tolonium Chloride
D024043:Collagen Type II; D018809:Proliferating Cell Nuclear Antigen; D014048:Tolonium Chloride