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Neuropeptide deficient mice have attenuated nociceptive, vascular, and inflammatory changes in a tibia fracture model of complex regional pain syndrome

Tian-Zhi Guo1, Tzuping Wei1, Xiaoyou Shi123, Wen-Wu Li123, Saiyun Hou1, Liping Wang1, Kazutake Tsujikawa4, Kenner C Rice5, Kejun Cheng5, David J Clark23 and Wade S Kingery1*

Author Affiliations

1 Physical Medicine and Rehabilitation Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA

2 Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA

3 Department of Anesthesiolgy, Stanford University School of Medicine, Stanford, CA, USA

4 Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan

5 Drug Design and Synthesis Section, Chemical Biology Research Branch, National Institute on Drug Abuse and National Institute on Alcohol Abuse and Alcoholism, NIH, Bethesda, MD, USA

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Molecular Pain 2012, 8:85  doi:10.1186/1744-8069-8-85

Published: 28 November 2012



Distal limb fracture in man can induce a complex regional pain syndrome (CRPS) with pain, warmth, edema, and cutaneous inflammation. In the present study substance P (SP, Tac1−/−) and CGRP receptor (RAMP1−/−) deficient mice were used to investigate the contribution of neuropeptide signaling to CRPS-like changes in a tibia fracture mouse model. Wildtype, Tac1−/−, and RAMP1−/− mice underwent tibia fracture and casting for 3 weeks, then the cast was removed and hindpaw mechanical allodynia, unweighting, warmth, and edema were tested over time. Hindpaw skin was collected at 3 weeks post-fracture for immunoassay and femurs were collected for micro-CT analysis.


Wildtype mice developed hindpaw allodynia, unweighting, warmth, and edema at 3 weeks post-fracture, but in the Tac1−/− fracture mice allodynia and unweighting were attenuated and there was no warmth and edema. RAMP1−/− fracture mice had a similar presentation, except there was no reduction in hindpaw edema. Hindpaw skin TNFα, IL-1β, IL-6 and NGF levels were up-regulated in wildtype fracture mice at 3 weeks post-fracture, but in the Tac1−/− and RAMP1−/− fracture mice only IL-6 was increased. The epidermal keratinocytes were the cellular source for these inflammatory mediators. An IL-6 receptor antagonist partially reversed post-fracture pain behaviors in wildtype mice.


In conclusion, both SP and CGRP are critical neuropeptide mediators for the pain behaviors, vascular abnormalities, and up-regulated innate immune responses observed in the fracture hindlimb. We postulate that the residual pain behaviors observed in the Tac1−/− and RAMP1−/− fracture mice are attributable to the increased IL-6 levels observed in the hindpaw skin after fracture.

Substance P; Calcitonin gene-related peptide; Fracture; Complex regional pain syndrome; Inflammation; Pain; Cytokine; Nerve growth factor