A Mouse Controlled Cortical Impact Model of Traumatic Brain Injury for Studying Blood–Brain Barrier Dysfunctions

  • Himakarnika Alluri
  • Chinchusha Anasooya Shaji
  • Matthew L. Davis
  • Binu TharakanEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1717)


Traumatic brain injury (TBI) is one of the leading causes of death and disability worldwide. It is a silently growing epidemic with multifaceted pathogenesis, and current standards of treatments aim to target only the symptoms of the primary injury, while there is a tremendous need to explore interventions that can halt the progression of the secondary injuries. The use of a reliable animal model to study and understand the various aspects the pathobiology of TBI is extremely important in therapeutic drug development against TBI-associated complications. The controlled cortical impact (CCI) model of TBI described here, uses a mechanical impactor to inflict a mechanical injury into the mouse brain. This method is a reliable and reproducible approach to inflict mild, moderate or severe injuries to the animal for studying TBI-associated blood–brain barrier (BBB) dysfunctions, neuronal injuries, brain edema, neurobehavioral changes, etc. The present method describes how the CCI model could be utilized for determining the BBB dysfunction and hyperpermeability associated with TBI. Blood–brain barrier disruption is a hallmark feature of the secondary injury that occur following TBI, frequently associated with leakage of fluid and proteins into the extravascular space leading to vasogenic edema and elevation of intracranial pressure. The method described here focuses on the development of a CCI-based mouse model of TBI followed by the evaluation of BBB integrity and permeability by intravital microscopy as well as Evans Blue extravasation assay.

Key words

Blood–brain barrier Controlled cortical impact Traumatic brain injury Intravital microscopy Evans Blue Edema Intracranial pressure Central nervous system Hyperpermeability Endothelial permeability 



The work presented in this chapter was supported by Scott & White Academic Operations Funds to Dr. Tharakan. The authors would like to apologize to investigators whose works are not cited in this methodological report due to space limitations and the personal perspective with which this chapter has been prepared.


  1. 1.
    Deaglio S, Robson SC (2011) Ectonucleotidases as regulators of purinergic signaling in thrombosis, inflammation, and immunity. Adv Pharmacol 61:301–332CrossRefPubMedGoogle Scholar
  2. 2.
    Shen Q, MH W, Yuan SY (2009) Endothelial contractile cytoskeleton and microvascular permeability. Cell Health Cytoskelet 1:43–50CrossRefGoogle Scholar
  3. 3.
    Kumar P, Shen Q, Pivetti CD, Lee ES, Wu MH, Yuan SY (2009) Molecular mechanisms of endothelial hyperpermeability: implications in inflammation. Expert Rev Mol Med 11:e19CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Yuan SY (2002) Protein kinase signaling in the modulation of microvascular permeability. Vasc Pharmacol 39:213–223CrossRefGoogle Scholar
  5. 5.
    Pearson WS, Sugerman DE, McGuire LC, Coronado VG (2012) Emergency department visits for traumatic brain injury in older adults in the United States: 2006-08. West J Emerg Med 13:289–293CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Alluri H, Wiggins-Dohlvik K, Davis ML, Huang JH, Tharakan B (2015) Blood-brain barrier dysfunction following traumatic brain injury. Metab Brain Dis 30(5):1093–1104CrossRefPubMedGoogle Scholar
  7. 7.
    Thurman DJ, Branche CM, Sniezek JE (1998) The epidemiology of sports-related traumatic brain injuries in the United States: recent developments. J Head Trauma Rehabil 3:1–8Google Scholar
  8. 8.
    Scott BN, Roberts DJ, Robertson HL, Kramer AH, Laupland KB, Ousman SS, Kubes P, Zygun DA (2013) Incidence, prevalence, and occurrence rate of infection among adults hospitalized after traumatic brain injury: study protocol for a systematic review and meta-analysis. Syst Rev 2:68CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Maas AIR, Stocchetti N, Bullock R (2008) Moderate and severe traumatic brain injury in adults. Lancet Neurol 7:728–741CrossRefPubMedGoogle Scholar
  10. 10.
    Parikh S, Koch M, Narayan RK (2007) Traumatic brain injury. Int Anesthesiol Clin 45:119–135CrossRefPubMedGoogle Scholar
  11. 11.
    O'Connor WT, Smyth A, Gilchrist MD (2011) Animal models of traumatic brain injury: a critical evaluation. Pharmacol Ther 130:106–113CrossRefPubMedGoogle Scholar
  12. 12.
    Kasper C, Yvette C (2015) Traumatic brain injury. In: Kasper C, Conley Y (eds) Annual review of nursing research, vol 33. Springer, New YorkGoogle Scholar
  13. 13.
    Gean AD, Fischbein NJ (2010) Head Trauma. Neuroimaging Clin N Am 20:527–556CrossRefPubMedGoogle Scholar
  14. 14.
    Davidoff G, Morris J, Roth E, Bleiberg J (1985) Closed head injury in spinal cord injured patients: retrospective study of loss of consciousness and post-traumatic amnesia. Arch Phys Med Rehabil 66:41–43PubMedGoogle Scholar
  15. 15.
    Cuccurullo S (2012) Physical medicine and rehabilitation pocket companion. Am J Phys Med Rehabil 91(8):727CrossRefGoogle Scholar
  16. 16.
    Xiong Y, Mahmood A, Chopp M (2013) Animal models of traumatic brain injury. Nat Rev Neurosci 14:128–142CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Nidavani RB, Mahalakshmi AM, Shalawadi M (2014) Vascular permeability and Evans blue dye: a physiological and pharmacological approach. J Appl Pharm Sci 4:106–113Google Scholar
  18. 18.
    Unterberg AW, Stover J, Kress B, Kiening KL (2004) Edema and brain trauma. Neuroscience 129:1021–1029CrossRefPubMedGoogle Scholar
  19. 19.
    Khan M, Im YB, Shunmugavel A, Gilg AG, Dhindsa RK, Singh AK, Singh IJ (2009) Administration of S-nitroglutathione after traumatic brain injury protects the neurovascular unit and reduces secondary injury in a rat model of controlled cortical impact. J Neuroinflammation 6:32CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Masedunskas A, Porat-Shliom N, Tora M, Milberg O, Weigert R (2013) Intravital microscopy for imaging subcellular structures in live mice expressing fluorescent proteins. J Vis Exp (79)Google Scholar
  21. 21.
    Marques PE, Oliveira AG, Amaral SS, Nunes-Silva A, Almeida AFS (2012) Intravital microscopy: taking a close look inside the living organisms. Afr J Microbiol Res 6:1603–1614Google Scholar
  22. 22.
    Taqueti VR, Jaffer FA (2013) High-resolution molecular imaging via intravital microscopy: illuminating vascular biology in vivo. Integr Biol (Camb) 5:278–290CrossRefGoogle Scholar
  23. 23.
    Radu M, Chernoff J (2013) An in vivo assay to test blood vessel permeability. J Vis Exp (73):e50062Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Himakarnika Alluri
    • 1
  • Chinchusha Anasooya Shaji
    • 1
  • Matthew L. Davis
    • 1
  • Binu Tharakan
    • 2
    Email author
  1. 1.Department of Surgery, Texas A&M University Health Science Center, College of MedicineBaylor Scott and White Research InstituteTempleUSA
  2. 2.Department of SurgeryTexas A&M University Health Science Center, College of Medicine, Baylor Scott and White Research InstituteTempleUSA

Personalised recommendations