Yonsei Med J.  2014 Jan;55(1):185-190. 10.3349/ymj.2014.55.1.185.

Transient Receptor Potential Vanilloid 1-Immunoreactive Innervation Increases in Fractured Rat Femur

• Kawarai Y
• Suzuki M
• Yoshino K
• Inoue G
• Orita S
• Yamauchi K
• Aoki Y
• Ishikawa T
• Miyagi M
• Kamoda H
• Kubota G
• Sakuma Y
• Oikawa Y
• Inage K
• Sainoh T
• Sato J
• Nakamura J
• Takaso M
• Toyone T
• Takahashi
• Ohtori S

• KMID: 1779904
• DOI: http://doi.org/10.3349/ymj.2014.55.1.185

Abstract

PURPOSE
Pain from vertebral or femoral neck fractures is a particularly important problem in clinical orthopaedics. Transient receptor potential vanilloid 1 (TRPV1) is a ligand-gated nonselective cation channel, and there are recent reports on an association between bone pain and TRPV1. However, an increase in TRPV1 activity has not been reported following femoral fracture.
MATERIALS AND METHODS
We applied a neurotracer [Fluoro-gold (FG)] onto femur to detect dorsal root ganglia (DRGs) innervating the cortex of the femur in 30 Sprague Dawley rats. Seven days after application, a closed mid-diaphyseal fracture of the femur was performed. FG labeled TRPV1-immunoreactive (ir) DRGs innervating the femur were examined in nonfractured controls, and 3 days, 1 week, 2 weeks, and 4 weeks after fracture. We evaluated bone healing of the femur and compared the ratio of TRPV1-ir DRG neurons innervating the femur at the time points.
RESULTS
Four weeks after fracture, complete bone union was observed. There was no significant difference in the ratio of FG labeled DRG neurons to total DRG neurons at each time point. The percentages of TRPV1-ir neurons in DRGs innervating the femur at 3 days and 1 week after fracture were significantly higher than those in control, 2 weeks, and 4 weeks after fracture (p<0.05).
CONCLUSION
Fracture induced an increase of TRPV1-ir neurons in DRGs innervating the fractured femur within 3 days, and decreased during bone healing over 4 weeks. These findings show that TRPV1 may play a role in sensory sensation of bone fracture pain.

Keyword

Fracture; pain; transient receptor potential vanilloid 1; femur; rat

MeSH Terms

Animals
Female
Femur/*innervation/*metabolism
Immunohistochemistry
Rats
Rats, Sprague-Dawley
TRPV Cation Channels/*metabolism
TRPV Cation Channels

Figure

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  Fig. 1 Radiographs showing at 3 days, 1 week, 2 weeks, and 4 weeks after fracture. (A) 3 days, (B) 1 week, (C) 2 weeks, and (D) 4 weeks.

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  Fig. 2 Innervation of the left femur at 2 weeks. FG-labeled (A) and TRPV1 immunoreactive DRG neurons (B) (A and B are the same sections. Arrows indicate double labeled neurons). FG, Fluoro-gold; TRPV1, transient receptor potential vanilloid 1; DRG, dorsal root ganglia.

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  Fig. 3 Distribution and ratio of FG-labeled DRG neurons innervating the left femur in control, 3 days, 1 week, 2 weeks, and 4 weeks groups. FG-labeled DRG neurons are present from L1 through L6, and there was no significant difference in the percentage of FG-labeled neurons of all DRG neurons among control, 3 days, 1 week, 2 weeks, and 4 weeks groups. FG, Fluoro-gold; DRG, dorsal root ganglion.

• 

  Fig. 4 Distribution and ratio of FG-labeled and TRPV1-ir DRG neurons innervating the left femur among control, 3 days, 1 week, 2 weeks, and 4 weeks groups. FG-labeled and TRPV1-ir DRG neurons innervating the left femur are present from L1 through L6. The percentages of TRPV1-ir neurons in the DRGs innervating the femur at 3 days and 1 week after fracture were significantly higher than those in nonfractured control, and 2 weeks and 4 weeks after fracture (p<0.05). FG, Fluoro-gold; TRPV1, transient receptor potential vanilloid 1; DRG, dorsal root ganglion; ir, immunoreactive.

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