In vivo model of Hirschsprung-associated enterocolitis using benzalkonium chloride

  • Iskandar Rahardjo Budianto Biomedical Doctoral Program, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; Department of Surgery, School of Medicine and Health Sciences, Universitas Katolik Indonesia Atma Jaya, Jakarta, Indonesia
  • Agus Firmansyah Department of Pediatric, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Yefta Moenadjat Department of Surgery, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-1109-2540
  • Ahmad Aulia Jusuf Department of Histology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia
  • Vivian Soetikno Department of Pharmacology and Therapeutic, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-2123-6448
Keywords: animal model, colitis, ganglions, Hirschsprung’s disease
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Abstract

BACKGROUND Hirschsprung-associated enterocolitis (HAEC) is a life-threatening complication of Hirschsprung’s disease. Studies using animal models on the pathogenesis of HAEC are limited. Thus, this study aimed to establish a rat model of HAEC using topical application of 0.1% benzalkonium chloride (BAC) in the sigmoid colon.

METHODS 55 male Sprague Dawley rats aged 10−12 weeks old were separated into 11 groups. The control group (n = 5) was euthanized on day-7, and the other 10 groups (n = 5 in each group) treated with 0.1% BAC in the sigmoid colon for 15 min to induce Hirschsprung’s disease were euthanized on day-7, -10, -12, -14, -17, -19, -21, -23, -25, and -28. The sigmoid colon was excised, fixed in formalin, and sectioned for histological examinations with hematoxylin and eosin staining. The degree of HAEC was compared within all groups.

RESULTS Rats that were sacrificed on day-7 to -12 showed the 1st degree or early HAEC, which was most likely caused by BAC application. The 2nd degree of HAEC occurred in rats that were sacrificed on day-14 that showed a macrophage infiltration in the sigmoid colon, thus fulfilled the initial criteria for HAEC (p = 0.0025 versus control). The degree of enterocolitis increased with time, and the highest degree was found in rats that were sacrificed on day-28 (p<0.001 versus control).

CONCLUSIONS Topical application of 0.1% BAC for 15 min was successfully produced HAEC model in rats, which was occurred on day-14 after the application. This model provides a useful resource for further research on the pathogenesis of HAEC.

Author Biography

Vivian Soetikno, Department of Pharmacology and Therapeutic, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia

SCOPUS h-index: 18

References

  1. Gosain A, Brinkman AS. Hirschsprung's associated enterocolitis. Curr Opin Pediatr. 2015;27(3):364-9. https://doi.org/10.1097/MOP.0000000000000210

  2. Teitelbaum DH, Coran AG. Enterocolitis. Semin Pediatr Surg. 1998;7(3):162-9. https://doi.org/10.1016/S1055-8586(98)70012-5

  3. Pini Prato A, Rossi V, Avanzini S, Mattioli G, Disma N, Jasonni V. Hirschsprung's disease: what about mortality? Pediatr Surg Int. 2011;27(5):473-8. https://doi.org/10.1007/s00383-010-2848-2

  4. Austin KM. The pathogenesis of Hirschsprung's disease-associated enterocolitis. Semin Pediatr Surg. 2012;21(4):319-27. https://doi.org/10.1053/j.sempedsurg.2012.07.006

  5. Frykman PK, Short SS. Hirschsprung-associated enterocolitis: prevention and therapy. Semin Pediatr Surg. 2012;21(4):328-35. https://doi.org/10.1053/j.sempedsurg.2012.07.007

  6. Yoneda A, Shima H, Nemeth L, Oue T, Puri P. Selective chemical ablation of the enteric plexus in mice. Pediatr Surg Int. 2002;18(4):234-7. https://doi.org/10.1007/s003830100681

  7. Garcia SB, Minto SB, de Souza Marques I, Kannen V. Myenteric denervation of the gut with benzalkonium chloride: a review of forty years of an experimental model. Can J Gastroenterol Hepatol. 2019;2019:3562492. https://doi.org/10.1155/2019/3562492

  8. Langness S, Kojima M, Coimbra R, Eliceiri BP, Costantini TW. Enteric glia cells are critical to limiting the intestinal inflammatory response after injury. Am J Physiol Gastrointest Liver Physiol. 2017;312(3):G274-82. https://doi.org/10.1152/ajpgi.00371.2016

  9. Qin HH, Lei N, Mendoza J, Dunn JC. Benzalkonium chloride-treated anorectums mimicked endothelin-3-deficient aganglionic anorectums on manometry. J Pediatr Surg. 2010;45(12):2408-11. https://doi.org/10.1016/j.jpedsurg.2010.08.045

  10. Moore SW, Zaahl MG. Tissue specific somatic mutations and aganglionosis in Hirschsprung's disease. J Pediatr Surg. 2014;49(2):258-61. https://doi.org/10.1016/j.jpedsurg.2013.11.035

  11. Wagner JP, Sullins VF, Khalil HA, Dunn JC. A durable model of Hirschsprung's colon. J Pediatr Surg. 2014;49(12):1804-8. https://doi.org/10.1016/j.jpedsurg.2014.09.024

  12. Teitelbaum DH, Caniano DA, Qualman SJ. The pathophysiology of Hirschsprung's-associated enterocolitis: importance of histologic correlates. J Pediatr Surg. 1989;24(12):1271-7. https://doi.org/10.1016/S0022-3468(89)80566-4

  13. Cheng Z, Dhall D, Zhao L, Wang HL, Doherty TM, Bresee C, et al. Murine model of Hirschsprung-associated enterocolitis. I: phenotypic characterization with development of a histopathologic grading system. J Pediatr Surg. 2010;45(3):475-82. https://doi.org/10.1016/j.jpedsurg.2009.06.009

  14. Yu H, Pan W, Wang H, Gao Y. A time-limited and partially reversible model of hypoganglionosis induced by benzalkonium chloride treatment. Neurochem Res. 2016;41(5):1138-44. https://doi.org/10.1007/s11064-015-1806-8

  15. Baudouin C, Labbe ́ A, Liang H, Pauly A, Brignole-Baudouin F. Preservatives in eyedrops: the good, the bad and the ugly. Prog Retin Eye Res. 2010;29(4):312-34. https://doi.org/10.1016/j.preteyeres.2010.03.001

  16. Michée S, Brignole-Baudouin F, Riancho L, Rostene W, Baudouin C, Labbé A. Effects of benzalkonium chloride on THP-1 differentiated macrophages in vitro. PLoS One. 2013;8(8):e72459. https://doi.org/10.1371/journal.pone.0072459

  17. 2 final report on the safety assessment of benzalkonium chloride. J Am Coll Toxicol. 1989;8(4):589-625. https://doi.org/10.3109/10915818909010524

  18. Heanue TA, Pachnis V. Enteric nervous system development and Hirschsprung's disease: advances in genetic and stem cell studies. Nat Rev Neurosci. 2007;8:466-79. https://doi.org/10.1038/nrn2137

Published
2021-08-27
How to Cite
1.
Budianto IR, Firmansyah A, Moenadjat Y, Jusuf AA, Soetikno V. <em>In vivo</em&gt; model of Hirschsprung-associated enterocolitis using benzalkonium chloride. Med J Indones [Internet]. 2021Aug.27 [cited 2021Dec.3];1(1). Available from: https://mji.ui.ac.id/journal/index.php/mji/article/view/5339
Section
Basic Medical Research