Yifei Huang1, Chunfang Jin1, Yun Xiang1, Lei Wang1, Zimeng Wang1, Wei Wang1, Yujuan He2. 1. Key Laboratory of Clinical Laboratory Diagnostics, Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. 2. Key Laboratory of Clinical Laboratory Diagnostics, Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China; Email: yujuanhe@cqmu.edu.cn.
Abstract
OBJECTIVE: To develop a mouse model for acute otitis media (AOM) via transbullar injection method and evaluate its feasibility and practicability. METHODS: The middle ears (ME) of C57BL/6 mice were inoculated via transbullar injection method with 5 µl streptococcus pneumoniae (S.pn) 19F suspension (1×10(7) CFU/ml), and the control group was inoculated equivalent phosphate buffered solution (PBS). Behavior changes were observed daily following inoculation. The ME tissues for histological examination and the middle ear lavage fluid (MELF) for total cells quantification, S.pn load determination and cytokines measurement were collected at 12 h, day 1, 2, 3, 5, 7 after inoculation, respectively. RESULTS: Within 24 hours after instillation, the density of S.pn and the level of acute inflammatory cytokines in ME cavity increased rapidly, some mucosal hyperplasia was evident and leukocytic infiltration (primarily neutrophils) began. The level of ME inflammatory response reached maximal at 2-3 days after inoculation, with extensive effusion, leukocytic infiltration and mucosal thickening. Meanwhile, the density of S.pn decreased gradually. Bacterial clearance was completed by day 5 with extensive resolution of ME inflammation, although mucosal hyperplasia did not resolute until day 7. CONCLUSION: A mouse model for AOM is successfully established via transbullar injection method, laying foundation for future study of AOM.
OBJECTIVE: To develop a mouse model for acute otitis media (AOM) via transbullar injection method and evaluate its feasibility and practicability. METHODS: The middle ears (ME) of C57BL/6 mice were inoculated via transbullar injection method with 5 µl streptococcus pneumoniae (S.pn) 19F suspension (1×10(7) CFU/ml), and the control group was inoculated equivalent phosphate buffered solution (PBS). Behavior changes were observed daily following inoculation. The ME tissues for histological examination and the middle ear lavage fluid (MELF) for total cells quantification, S.pn load determination and cytokines measurement were collected at 12 h, day 1, 2, 3, 5, 7 after inoculation, respectively. RESULTS: Within 24 hours after instillation, the density of S.pn and the level of acute inflammatory cytokines in ME cavity increased rapidly, some mucosal hyperplasia was evident and leukocytic infiltration (primarily neutrophils) began. The level of ME inflammatory response reached maximal at 2-3 days after inoculation, with extensive effusion, leukocytic infiltration and mucosal thickening. Meanwhile, the density of S.pn decreased gradually. Bacterial clearance was completed by day 5 with extensive resolution of ME inflammation, although mucosal hyperplasia did not resolute until day 7. CONCLUSION: A mouse model for AOM is successfully established via transbullar injection method, laying foundation for future study of AOM.