In vivo model of Hirschsprung-associated enterocolitis using benzalkonium chloride
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.
Gosain A, Brinkman AS. Hirschsprung's associated enterocolitis. Curr Opin Pediatr. 2015;27(3):364-9. https://doi.org/10.1097/MOP.0000000000000210
Teitelbaum DH, Coran AG. Enterocolitis. Semin Pediatr Surg. 1998;7(3):162-9. https://doi.org/10.1016/S1055-8586(98)70012-5
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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