Anti-inflammatory treatment increases angiogenesis during early fracture healing

ObjectivesBoth inflammation and angiogenesis are crucial for normal fracture healing. The goal of this work was to determine how anti-inflammatory treatment affects angiogenesis during early stages of fracture repair.MethodsTibia fractures were created in adult mice and animals were treated with indomethacin (2 mg/kg/day), a non-steroidal anti-inflammatory drug, or PBS once a day beginning from 1 day before fracture and continuing to 6 days after fracture. Animals were killed at 7, 14, and 28 days after injury for histomorphometric analysis of fracture healing. A second group of animals were killed at 3 and 7 days after injury to measure tissue levels of VEGF and interleukin-1 beta (IL-1β). A third group of animals were killed at 3 and 7 days after injury for stereology analysis of macrophage and neutrophil infiltration and tissue vascularization.ResultsIndomethacin significantly decreased bone and cartilage formation at 7 days after fracture compared to controls. Indomethacin decreased the tissue levels of IL-1β at 3 days after fracture but did not affect the recruitment of macrophages or neutrophils to injured limbs. Indomethacin-treated fractures had similar length density and surface density of vasculature as the controls at 3 days after injury. At 7 days after fracture, vasculature in indomethacin-treated fractures exhibited higher length density and surface density than that in controls. By 28 days after injury, indomethacin-treated fractures still exhibited defects in fracture repair.ConclusionsAnti-inflammatory treatments using indomethacin impair bone and cartilage formation and increase tissue vascularization in the callus during early fracture healing.

[1]  E. Sudmann,et al.  Effect of indomethacin on fracture healing in rats. , 1976, Acta orthopaedica Scandinavica.

[2]  E. Sudmann,et al.  Inhibition of fracture healing by indomethacin in rats , 1979, European journal of clinical investigation.

[3]  A. Wase,et al.  Indomethacin and aspirin: effect of nonsteroidal anti-inflammatory agents on the rate of fracture repair in the rat. , 1980, Acta orthopaedica Scandinavica.

[4]  M. Elves,et al.  The effect of indomethacin upon experimental fractures in the rat. , 1982, Acta orthopaedica Scandinavica.

[5]  D. Faller,et al.  Interleukin 1 induces cultured human endothelial cell production of granulocyte-macrophage colony-stimulating factor. , 1987, The Journal of clinical investigation.

[6]  E. Sudmann,et al.  Fracture healing in rats inhibited by locally administered indomethacin. , 1992, Acta orthopaedica Scandinavica.

[7]  K Banovac,et al.  Effect of Nonsteroidal Antiinflammatory Drugs on Fracture Healing: A Laboratory Study in Rats , 1995, Journal of orthopaedic trauma.

[8]  T A Einhorn,et al.  The expression of cytokine activity by fracture callus , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[9]  M. Devidas,et al.  Production of interleukin-10 in human fracture soft-tissue hematomas. , 1996, Shock.

[10]  V. Howard,et al.  Unbiased Stereology: Three-Dimensional Measurement in Microscopy , 1998 .

[11]  John R. Johnson,et al.  The Effect of Postoperative Nonsteroidal Anti‐inflammatory Drug Administration on Spinal Fusion , 1998, Spine.

[12]  D. Macdonald,et al.  Nonunion of the femoral diaphysis. The influence of reaming and non-steroidal anti-inflammatory drugs. , 2000, The Journal of bone and joint surgery. British volume.

[13]  M. Schaffler,et al.  Prevention of fracture healing in rats by an inhibitor of angiogenesis. , 2001, Bone.

[14]  E. Schwarz,et al.  Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair. , 2002, The Journal of clinical investigation.

[15]  Diane Hu,et al.  A model for intramembranous ossification during fracture healing , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  J. Anglen,et al.  Heterotopic ossification prophylaxis with indomethacin increases the risk of long-bone nonunion. , 2003, The Journal of bone and joint surgery. British volume.

[17]  Thomas A Einhorn,et al.  Effects of the local mechanical environment on vertebrate tissue differentiation during repair: does repair recapitulate development? , 2003, Journal of Experimental Biology.

[18]  H. Donahue,et al.  Effect of COX-2-specific inhibition on fracture-healing in the rat femur. , 2004, The Journal of bone and joint surgery. American volume.

[19]  S. Reuben,et al.  High dose nonsteroidal anti-inflammatory drugs compromise spinal fusion , 2005, Canadian journal of anaesthesia = Journal canadien d'anesthesie.

[20]  K. Sairyo,et al.  Cyclooxygenase-2 inhibitor delays fracture healing in rats , 2005, Acta orthopaedica.

[21]  Diane Hu,et al.  Cellular basis for age‐related changes in fracture repair , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[22]  B. Goldberg,et al.  Effects of a cyclooxygenase-2 inhibitor (rofecoxib) on bone healing. , 2006, American journal of orthopedics.

[23]  Theodore Miclau,et al.  Assessing angiogenesis during fracture healing. , 2006, The Iowa orthopaedic journal.

[24]  P. Weinhold,et al.  Effect of COX-2 inhibitors and non-steroidal anti-inflammatory drugs on a mouse fracture model. , 2006, Injury.

[25]  M. Swiontkowski.  Nonsteroidal Antiinflammatory Drugs and Nonunion of Humeral Shaft Fractures , 2006 .

[26]  Gang Li,et al.  Nonsteroidal anti-inflammatory drug-induced fracture nonunion: an inhibition of angiogenesis? , 2006, The Journal of bone and joint surgery. American volume.

[27]  Subburaman Mohan,et al.  Microarray analysis of gene expression during the inflammation and endochondral bone formation stages of rat femur fracture repair. , 2006, Bone.

[28]  A. M. Simon,et al.  Dose and time-dependent effects of cyclooxygenase-2 inhibition on fracture-healing. , 2007, The Journal of bone and joint surgery. American volume.

[29]  D. Noonan,et al.  Inflammation, inflammatory cells and angiogenesis: decisions and indecisions , 2008, Cancer and Metastasis Reviews.

[30]  T. Karachalios,et al.  The effects of the short-term administration of low therapeutic doses of anti-COX-2 agents on the healing of fractures. An experimental study in rabbits. , 2007, The Journal of bone and joint surgery. British volume.

[31]  P. Dockery,et al.  The quantification of vascular beds: a stereological approach. , 2007, Experimental and molecular pathology.

[32]  E. Krall,et al.  Selective and nonselective cyclooxygenase-2 inhibitors and experimental fracture-healing. Reversibility of effects after short-term treatment. , 2007, The Journal of bone and joint surgery. American volume.

[33]  J. O'Connor,et al.  Celecoxib, NSAIDs and the skeleton. , 2008, Drugs of today.

[34]  S. Donnini,et al.  Prostaglandin E2 Regulates Angiogenesis via Activation of Fibroblast Growth Factor Receptor-1* , 2008, Journal of Biological Chemistry.

[35]  D. Hu,et al.  Effect of age on vascularization during fracture repair , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[36]  Thomas Pufe,et al.  The systemic angiogenic response during bone healing , 2009, Archives of Orthopaedic and Trauma Surgery.

[37]  B. Biglari,et al.  Clinical application of BMP 7 in long bone non-unions , 2009, Archives of Orthopaedic and Trauma Surgery.

[38]  Hai Yao,et al.  Regeneration of the articular surface of the rabbit synovial joint by cell homing: a proof of concept study , 2010, The Lancet.

[39]  D. Hu,et al.  Action of IL‐1β during fracture healing , 2010, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[40]  W. Olszewski,et al.  Genetic factors responsible for long bone fractures non-union , 2010, Archives of Orthopaedic and Trauma Surgery.

[41]  C. Colnot,et al.  Mechanical Stability Affects Angiogenesis During Early Fracture Healing , 2011, Journal of orthopaedic trauma.