A refinement approach in a mouse model of rehabilitation research. Analgesia strategy, reduction approach and infrared thermography in spinal cord injury

Autoři: Veronica Redaelli aff001;  Simonetta Papa aff002;  Gerardo Marsella aff003;  Giuliano Grignaschi aff003;  Alice Bosi aff004;  Nicola Ludwig aff005;  Fabio Luzi aff001;  Irma Vismara aff002;  Stefano Rimondo aff002;  Pietro Veglianese aff002;  Svetlana Tepteva aff001;  Silvia Mazzola aff001;  Pietro Zerbi aff006;  Luca Porcu aff007;  John V. Roughan aff008;  Gianfranco Parati aff004;  Laura Calvillo aff004
Působiště autorů: Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Milan, Italy aff001;  Department of Neuroscience. Laboratory of Biology of Neurodegenerative Disorders, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy aff002;  Animal Care Unit, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy aff003;  Department of Cardiovascular, Neural and Metabolic Sciences, S.Luca Hospital, Istituto Auxologico Italiano IRCCS, Milan, Italy aff004;  Dipartimento di Fisica, Università degli Studi di Milano, Milan, Italy aff005;  Dipartimento di Scienze Biomediche e Cliniche "L. Sacco", Università degli Studi di Milano, Milan, Italy aff006;  Department of Oncology, Laboratory of Methodology for Clinical Research, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy aff007;  Institute of Neuroscience, Comparative Biology Centre, Newcastle University, Newcastle upon Tyne, United Kingdom aff008;  Department of Medicine and Surgery, University of Milano-Bicocca, Milan, Italy aff009
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0224337


The principles of Refinement, Replacement and Reduction (3R’s) should be taken into account when animals must be used for scientific purpose. Here, a Reduction / Refinement approach was applied to the procedure of spinal cord injury (SCI), an animal model used in rehabilitation medicine research, in order to improve the quality of experiments, avoiding unnecessary suffering. The aims of this investigation were 1- to assess acute surgical pain in mice subjected to SCI, 2- to compare the efficacy of commonly used analgesia (three buprenorphine subcutaneous injection in 48 hours, 0,15 mg/kg each) with a combination of opioid and NSAID (one subcutaneous injection of 5 mg/kg carprofen before surgery followed by three buprenorphine subcutaneous injection in 48 hours, 0,15 mg/kg each) and 3- to test if Infrared Thermography (IRT) could be a potential new Refinement method to easily assess thermoregulation, an important metabolic parameter. Finally, we aimed to achieve these goals without recruiting animals on purpose, but using mice already scheduled for studies on SCI. By using behaviours analysis, we found that, despite being commonly used, buprenorphine does not completely relieve acute surgical pain, whereas the combination of buprenorphine and carprofen significantly decreases pain signs by 80%. IRT technology turned out to be a very useful Refinement tool being a non invasive methods to measure animal temperature, particularly useful when rectal probe cannot be used, as in the case of SCI. We could find that temperatures constantly and significantly increased until 7 days after surgery and then slowly decreased and, finally, we could observe that in the buprenorphine and carprofen treated group, temperatures were statistically lower than in the buprenorphine-alone treated mice. To our knowledge this is the first work providing an analgesic Refinement and a description of thermoregulatory response using the IRT technology, in mice subjected to SCI.

Klíčová slova:

Analgesia – Analgesics – Animal behavior – Ears – Mice – Mouse models – Spinal cord injury – Surgical and invasive medical procedures


1. Ciuffreda MC, Tolva V, Casana R, Gnecchi M, Vanoli E, Spazzolini C, et al. Rat experimental model of myocardial ischemia/reperfusion injury: An ethical approach to set up the analgesic management of acute post-surgical pain. PLoS One. 2014;9(4):e95913. doi: 10.1371/journal.pone.0095913 24756074

2. World Health Organization. Spinal cord injury [Internet]. 2013. http://www.who.int/news-room/fact-sheets/detail/spinal-cord-injury

3. Balcombe JP, Barnard ND, Sandusky C. Laboratory routines cause animal stress. Contemp Top Lab Anim Sci. 2004;43(6):42–51. 15669134

4. Lofgren J, Miller AL, Lee CCS, Bradshaw C, Flecknell P, Roughan J. Analgesics promote welfare and sustain tumour growth in orthotopic 4T1 and B16 mouse cancer models. Lab Anim. 2018;52(4):351–64. doi: 10.1177/0023677217739934 29207902

5. Musterman M (University of BC. Rat and mouse anasthesia and analgesia formulary and general drug information. 2016;1–14. https://animalcare.ubc.ca/sites/default/files/documents/Guideline-Rodent%20Anesthesia%20Analgesia%20Formulary%282016%29.pdf

6. Forgione N, Chamankhah M, Fehlings MG. A Mouse Model of Bilateral Cervical Contusion-Compression Spinal Cord Injury. J Neurotrauma. 2016;34(6):1227–39.

7. Brommer B, Engel O, Kopp MA, Watzlawick R, Müller S, Prüss H, et al. Spinal cord injury-induced immune deficiency syndrome enhances infection susceptibility dependent on lesion level. Brain. 2016;139(3):692–707.

8. Prüss H, Tedeschi A, Thiriot A, Lynch L, Loughhead SM, Stutte S, et al. Spinal cord injury-induced immunodeficiency is mediated by a sympathetic-neuroendocrine adrenal reflex. Nat Neurosci. 2017;20(11):1549–59. doi: 10.1038/nn.4643 28920935

9. Yang L, Ge D, Chen X, Jiang C, Zheng S. MiRNA-544a regulates the inflammation of spinal cord injury by inhibiting the expression of NEUROD4. Cell Physiol Biochem. 2018;51(4):1921–31. doi: 10.1159/000495717 30513512

10. Saini V, Loers G, Kaur G, Schachner M, Jakovcevski I. Impact of neural cell adhesion molecule deletion on regeneration after mouse spinal cord injury. Eur J Neurosci. 2016;44(1):1734–46. doi: 10.1111/ejn.13271 27178448

11. Lee P, Ho KKY, Lee P, Greenfield JR, Ho KKY, Greenfield JR. Hot fat in a cool man: Infrared thermography and brown adipose tissue. Diabetes, Obes Metab. 2011;13(1):92–3.

12. Society of Nuclear Medicine (1953-) J, Knowles S, Taschereau R, Chatziioannou A, Stout D. Murine brown fat metabolism measured by infrared thermography correlates with PET FDG uptake [Internet]. Vol. 54, Journal of Nuclear Medicine. Society of Nuclear Medicine; 2013 May [cited 2019 Jul 23]. http://jnm.snmjournals.org/cgi/content/short/54/supplement_2/1120

13. Ferreira V, Francisco N, Belloni M, Aguirre G, Caldara F, Nääs I, et al. Infrared thermography applied to the evaluation of metabolic heat loss of chicks fed with different energy densities. Rev Bras Ciência Avícola. 2011;13(2):113–8.

14. Su TY, Ho WT, Chiang SC, Lu CY, Chiang HK, Chang SW. Infrared thermography in the evaluation of meibomian gland dysfunction. J Formos Med Assoc [Internet]. 2017;116(7):554–9. http://dx.doi.org/10.1016/j.jfma.2016.09.012 27817990

15. Rekant SI. Veterinary applications of infrared thermography. Am J Vet Res. 2016;77(1):98–107. doi: 10.2460/ajvr.77.1.98 26709943

16. Vianna D.M.L. and Carrive P. Changes in cutaneous and body temperature during and after conditioned fear to context in the rat. Proc Aust Neurosci Soc. 2005;21:2505–2512.

17. Ludwig N, Gargano M, Luzi F, Carenzi C, Verga M. Technical note: Applicability of infrared thermography as a non invasive measurement of stress in rabbit. World Rabbit Sci. 2007;15(4):199–205.

18. Fornasier M., Redaelli V., Tarantino A., Luzi F. VM. Infrared thermography (IRT) in nude mice: an alternative method for body temperature measurement. In: Atti Scand FELASA 2010, Helsinki, June 14–17, 2010. 2010.

19. Vianna D.M.L. and Carrive P. Changes in cutaneous and body temperature during and after conditioned fear to context in the rat. Eur J Neurosci. 2005;21:2505–12. doi: 10.1111/j.1460-9568.2005.04073.x 15932607

20. Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG. The ARRIVE guidelines Animal Research: Reporting In Vivo Experiments. 2010;(June):1–2.

21. Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman and DG. The ARRIVE Guidelines Checklist. https://www.nc3rs.org.uk/arrive-guidelines [Internet]. 2010; https://www.nc3rs.org.uk/sites/default/files/documents/Guidelines/NC3Rs%20ARRIVE%20Guidelines%20Checklist%28fillable%29.pdf

22. Stokes EL, Flecknell PA, Richardson CA. Reported analgesic and anaesthetic administration to rodents undergoing experimental surgical procedures. Lab Anim. 2009;43(2):149–54. doi: 10.1258/la.2008.008020 19116297

23. Basso DM, Fisher LC, Anderson AJ, Jakeman LB, McTigue DM, Popovich PG. Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains. J Neurotrauma. 2006;23(5):635–59. doi: 10.1089/neu.2006.23.635 16689667

24. Roughan J V., Flecknell PA. Behavioural effects of laparotomy and analgesic effects of ketoprofen and carprofen in rats. Pain. 2001;90(1–2):65–74. doi: 10.1016/s0304-3959(00)00387-0 11166971

25. Langford DJ, Bailey AL, Chanda ML, Clarke SE, Drummond TE, Echols S, et al. Coding of facial expressions of pain in the laboratory mouse. Nat Methods. 2010;7(6):447–9. doi: 10.1038/nmeth.1455 20453868

26. Keating SCJ, Thomas AA, Flecknell PA, Leach MC. Evaluation of EMLA Cream for Preventing Pain during Tattooing of Rabbits: Changes in Physiological, Behavioural and Facial Expression Responses. PLoS One. 2012;7(9):e44437. doi: 10.1371/journal.pone.0044437 22970216

27. Ren K, Dubner R. Interactions between the immune and nervous systems in pain. Nat Med [Internet]. 2010;16(11):1267–76. http://www.ncbi.nlm.nih.gov/pubmed/20948535 doi: 10.1038/nm.2234

28. Wang H, Sun H, Della Penna K, Benz RJ, Xu J, Gerhold DL, et al. Chronic neuropathic pain is accompanied by global changes in gene expression and shares pathobiology with neurodegenerative diseases. Neuroscience. 2002;114(3):529–46. doi: 10.1016/s0306-4522(02)00341-x 12220557

29. Gemes G., Rigaud M. Dean C., Hopp F.A., Hogan Q.H., Seagard J. Baroreceptor Reflex is Suppressed in Rats that Develop Hyperalgesia Behavior after Nerve Injury. Pain 2009 December; 146(3):293–300. doi: 10.1016/j.pain.2009.07.040 19729245

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2019 Číslo 10
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