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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

The tibial fracture-pin model is a clinically relevant model of orthopedic trauma comprising a unilateral open tibial fracture with intramedullary nail internal fixation and simultaneous injury to the tibialis anterior muscle. Thermal sensitivity in this model can be measured using a 45 s hot plate paradigm.

Abstract

The tibial fracture-pin model is a mouse model of orthopedic trauma and surgery that recapitulates the complex muscle, bone, nerve, and connective tissue damage that manifests with this type of injury in humans. This model was developed because previous models of orthopedic trauma did not include simultaneous injury to multiple tissue types (bone, muscle, nerves) and were not truly representative of human complex orthopedic trauma. The authors therefore modified previous models of orthopedic trauma and developed the tibial fracture-pin model. This modified fracture model consists of a unilateral open tibial fracture with intramedullary nail (IMN) internal fixation and simultaneous tibialis anterior (TA) muscle injury, resulting in mechanical allodynia that lasts up to 5 weeks post injury. This series of protocols outlines the detailed steps to perform the clinically relevant orthopedic trauma tibial fracture-pin model, followed by a modified hot plate assay to examine nociceptive changes after orthopedic injury. Taken together, these detailed, reproducible protocols will allow pain researchers to expand their toolkit for studying orthopedic trauma-induced pain.

Introduction

Orthopedic trauma accounts for 25% of all injuries sustained by nearly 500 million people each year worldwide1,2,3. Orthopedic trauma can be associated with complex muscle, bone, nerve, and connective tissue damage, necessitating hospitalization and surgery to ensure adequate recovery3,4. Acute and chronic pain after orthopedic trauma can result in significant physical, psychological, and financial burdens that affect a patient's quality of life1,4. Additi....

Protocol

All methods used while conducting this research were performed in compliance and with approval by the Stanford University Administrative Panel on Laboratory Animal Care (APLAC #33114) in accordance with American Veterinary Medical Association guidelines and the International Association for the Study of Pain. Mice (C57BL/6J, 9-11 weeks old upon arrival, 11-12 weeks old at study initiation) were housed 2-5 per cage and maintained on a 12 h light/dark cycle in a temperature-controlled environment with ad libitum a.......

Representative Results

The tibial fracture-pin orthotrauma model reproduces the bone, muscle, and pain-like behaviors seen in complex human injury. As shown in Figure 1C, the tibial fracture heals over time, forming a callus at the fracture site that is still seen at 4 weeks post injury. As a result of the lateral approach with the bone saw described above (step 3.5), the tibialis anterior muscle is injured, becoming extensively fibrotic, as seen by increased collagen deposition throughout the tissue (

Discussion

Critical steps within the protocol
It is crucial to maintain sterile conditions throughout the surgery. Moreover, proper animal care before, during, and after the surgery is paramount for the successful generation of the model. As mentioned earlier in the protocol, when performing the surgery, fracture the bone from the lateral side to ensure muscle injury. Take care not to fracture the tibia too low (below the advised junction between the middle and distal thirds of the tibia) because this will af.......

Acknowledgements

GM is supported by an NDSEG Graduate Fellowship and a Stanford Bio-X Honorary Graduate Fellowship. VLT is supported by NIH NIGMS grant #GM137906 and the Rita Allen Foundation.

....

Materials

NameCompanyCatalog NumberComments
27 G needlesMedsitis305136https://medsitis.com/products/bd-precisionglide-27-g-x-1-1-4-hypodermic-needles-305136?variant=39724583299
5-0 sutureesutureSN5668https://www.esutures.com/product/0-in-date/2-/132-/16552-medtronic-monosof-black-18-p-11-cutting-SN5668/
Alcohol swabsAmazonB00VS4F91Whttps://www.amazon.com/Dynarex-Alcohol-Prep-Sterile-Medium/dp/B00VS4F91W
Alternative drill bitsRio Grande341602https://www.riogrande.com/product/BuschTungstenVanadiumRoundBur
Set0314mm/341602
Bone saw drill attachmentAmazonB07DSXR3NYhttps://www.amazon.com/dp/B07DSXR3NY
BuprenorphineFidelis Pharmaceuticalshttps://ethiqaxr.com/ordering/
Ceramic implant (alternative to pin)RISystemRIS.221.103https://risystem.com/platefixation/mousescrew
Chux (Absorbent Underpad)Fisher ScientificNC0059881https://www.fishersci.com/shop/products/underpad-17x24-chux-300-cs/nc0059881#?keyword=true
C57BL/6J miceThe Jackson LaboratoryJax #00664https://www.jax.org/strain/000664
Cotton swabsUlineS-18991https://www.uline.com/Product/Detail/S-18991/First-Aid/Cotton-Tipped-Applicators-Industrial-6
Cutting pliersAmazonB076XYVS6Yhttps://www.amazon.com/iExcell-Diagonal-Cutting-Nippers-Chrome-Vanadium/dp/B076XYVS6Y
DrillChewy129044https://www.chewy.com/dremel-cordless-dog-cat-rotary-nail/dp/156127
Drill bitsAmazonB00HVIGSX2https://www.amazon.com/Universal-Diamond-Dremel-Rotary-Tool/dp/B00HVIGSX2
Electric shaverKent ScientificCL9990-KIThttps://www.kentscientific.com/products/trimmer-combo-kit/
Eye lubeAmazonB07H2NLCX5https://www.amazon.com/OptixCare-Lube-Plus-Hyaluron-Horses/dp/B07H2NLCX5
Gauze pads 2" x 2"AmazonB07GHDTB53https://www.amazon.com/Covidien-Curity-Sterile-Peel-Back-Package/dp/B07GHDTB53
Gauze pads 4" x 4"AmazonB00KJ6YFTChttps://www.amazon.com/Covidien-6309-Curity-Gauze-Pads/dp/B00KJ6YFTC
High definition video cameraThe Imaging SourceDFK 22AUC03https://www.theimagingsource.com/products/industrial-cameras/usb-2.0-color/dfk22auc03/?adsdyn&gclid=Cj0KCQiA3-yQBhD3ARIsAHuHT64uIIlImBvh_
toh-3GFSgBcL_fRc1gQTDyXlqDEa
Qu4n2_VbWEiRuIaAiueEALw_wcB
Inhalational anesthesia systemKent Scientifichttps://www.kentscientific.com/products/vaporizer-with-vetflo-single-channel-anesthesia-stand/
Iodine solutionAmazonB005FR7XIKhttps://www.amazon.com/Dynarex-Povidone-Iodine-Scrub-Solution/dp/B005FR7XIK
Iodine swab sticksAmazonB001V9QKMGhttps://www.amazon.com/POVIDONE-IODINE-SWAB-1202-25Box/dp/B001V9QKMG
IsofluraneCalifornia pet pharmacyhttps://www.californiapetpharmacy.com/fluriso-isoflurane-250ml.html
NCH Prism Softwarehttps://www.nchsoftware.com/prism/index.html
Plastic CylinderAmazonB08R5KM5B6https://www.amazon.com/FixtureDisplays-Acrylic-Diameter-Thickness-15140-8-NPF/dp/B08R5KM5B6
SalineFisher ScientificNC9054335https://www.fishersci.com/shop/products/saline-injection-0-9-10ml/NC9054335
ScalpelFisher Scientific12-000-162https://www.fishersci.com/shop/products/high-precision-10-style-scalpel-blade/12000162#?keyword=
Scalpel handleAmazonB0056ZX1R8https://www.amazon.com/Swann-Morton-Scalpel-Handle-blades/dp/B0056ZX1R8
Thermal place preference apparatusBIOSEBBIO-T2CThttps://www.bioseb.com/en/pain-thermal-allodynia-hyperalgesia/897-thermal-place-preference-2-temperatures-choice-nociception-test.html

References

  1. Edgley, C., et al. Severe acute pain and persistent post-surgical pain in orthopaedic trauma patients: a cohort study. British Journal of Anaesthesia. 123 (3), 350-359 (2019).
  2. Pointer, S. Trend....

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Tibial FractureOrthopedic InjuryMouse ModelTraumaBoneMuscleNerveConnective TissueIntramedullary NailTibialis Anterior MuscleMechanical AllodyniaNociceptive ChangesPain Research

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